CXCL10/IP-10: A Missing Link Between Inflammation and Anti- Angiogenesis in Preeclampsia?
By Gotsch, Francesca Romero, Roberto; Friel, Lara; Kusanovic, Juan Pedro; Espinoza, Jimmy; Erez, Offer; Than, Nandor Gabor; Mittal, Pooja; Edwin, Samuel; Yoon, Bo Hyun; Kim, Chong Jai; Mazaki-Tovi, Shali; Chaiworapongsa, Tinnakorn; Hassan, Sonia S
Abstract Objective. Interferon (IFN)-gamma inducible protein, CXCL10/IP-10, is a member of the CXC chemokine family with proinflammatory and anti-angiogenic properties. This chemokine has been proposed to be a key link between inflammation and angiogenesis. The aim of this study was to determine whether preeclampsia and delivery of a small for gestational age (SGA) neonate are associated with changes in maternal serum concentration of CXCL10/IP-10.
Study design. This cross-sectional study included patients in the following groups: (1) non-pregnant women (N = 49); (2) women with normal pregnancies (N = 89); (3) patients with preeclampsia (N = 100); and (4) patients who delivered an SGA neonate (N = 78). SGA was defined as birth weight below the 10th percentile. Maternal serum concentrations of CXCL10/IP-10 were measured by sensitive immunoassay. Non-parametric statistics were used for analysis.
Results. (1) Patients with normal pregnancies had a significantly higher median serum concentration of CXCL10/IP-10 than non-pregnant women (median 116.1 pg/mL, range 40.7-1314.3 vs. median 90.3 pg/mL, range 49.2-214.7, respectively; p = 0.002); (2) no significant correlation was found between maternal serum concentration of CXCL10/ IP-10 and gestational age (between 19 and 38 weeks); (3) there were no differences in median serum CXCL10/IP-10 concentrations between patients who delivered an SGA neonate and those with normal pregnancies (median 122.4 pg/mL, range 37.3-693.5 vs. median 116.1 pg/mL, range 40.7-1314.3, respectively; p > 0.05); (4) patients with preeclampsia had a higher median serum concentration of CXCL10/IP- 10 than normal pregnant women (median 156.4 pg/mL, range 47.4-645.9 vs. median 116.1 pg/mL, range 40.7-1314.3, respectively; p < 0.05); (5) patients with preeclampsia had a higher median concentration of CXCL10/IP-10 than those who delivered an SGA neonate (median 156.4 pg/mL, range 47.4-645.9 vs. median 122.4 pg/mL, range 37.3-693.5, respectively; p < 0.05).
Conclusions. Patients with preeclampsia have significantly higher serum concentrations of CXCL10/IP-10 than both normal pregnant women and mothers who have SGA neonates. These results are likely to reflect an anti-angiogenic state as well as an enhanced systemic inflammatory response in patients with preeclampsia. Alternatively, since preeclampsia and SGA share several mechanisms of disease, it is possible that a higher concentration of this chemokine may contribute to the clinical presentation of preeclampsia in patients with a similar intrauterine insult.
Keywords: Pregnancy, CXCL10, IP-10, chemokine, chemotactic cytokine, small for gestational age, SGA, angiogenesis
Introduction
Patients with preeclampsia and those with a small for gestational age (SGA) neonate share a number of pathophysiological characteristics including: (1) abnormal physiologic transformation of the spiral arteries [1-8]; (2) chronic uteroplacental ischemia [9- 16]; (3) endothelial cell dysfunction [17-24]; (4) an anti- angiogenic state [25-32]; and (5) intravascular inflammation [18,33- 37]. Furthermore, a biased Th1/Th2 (T-helper 1/T-helper 2) balance towards a Th1 response has been reported in preeclampsia [38-46] and SGA [47].
The human interferon-inducible protein 10 (IP-10 or CXCL10) is a chemokine of the CXC family [48]. A unique feature of members of this chemokine family is that they have pro-inflammatory properties and act as modulators of angiogenesis in conditions such as wound healing, ischemia, and neoplasia. These dual properties are related to the shared expression of specific chemokine receptors by leukocytes and endothelial cells [49-60].
IP-10 is inducible by pro-inflammatory stimuli such as interferon- gamma (IFN-gamma) [61-72], tumor necrosis factor-alpha (TNF-alpha) [70,73-79], viruses, and microbial products [66,70,80-85], directly or through activation of nuclear factor-kappa B (NF-kB) [81,82,86- 89]. It has been proposed that this chemokine is also involved in recruitment and potentiation of Th1 responses as well as in the pathogenesis of allograft rejection [90-103], multiple sclerosis [104-108], diabetes mellitus type 1 [109,110], Graves’ disease [111- 114], autoimmune thyroiditis [115,116], pulmonary fibrosis [117- 119], and cardiovascular diseases such as atherosclerosis [120] and coronary syndromes [121,122]. Importantly, IP-10 has potent anti- angiogenic activity, both in vitro and in vivo [123-126].
The balance between angiogenic and antiangiogenic (angiostatic) factors controlled by molecules involved in inflammatory processes may have an impact on the pathogenesis of many diseases [54,58].
We propose that IP-10 is involved in the pathophysiology of preeclampsia because this condition is characterized by intravascular inflammation and an anti-angiogenic state. The objective of this study was to compare the maternal serum IP-10 concentrations in normal pregnancy, preeclampsia, and SGA.
Methods
Study design
This retrospective cross-sectional study included patients with preeclampsia (N = 100), women who delivered an SGA neonate (N = 78), normal pregnant women (N = 89), and non-pregnant women (N = 49). All patients were enrolled at Hutzel Hospital, Detroit, MI, USA. All women provided written informed consent for the collection of clinical data and biological materials under protocols approved by the Institutional Review Boards of both Wayne State University and the National Institute of Child Health and Human Development of the National Institute of Health (NIH/DHHS). Many of these samples have been previously employed to study the biology of inflammation, hemostasis, angiogenesis regulation, and growth factor concentrations in non-pregnant women, normal pregnant women, and those with pregnancy complications.
Preeclampsia was defined in the presence of hypertension (systolic blood pressure >/=140 mmHg or diastolic blood pressure >/ =90 mmHg on at least two occasions, 4 hours to 1 week apart, after the 20th week of gestation) and proteinuria (>/=300 mg in a 24-hour urine collection, or two random urine specimens obtained 4 hours to 1 week apart containing >/=1+ protein by dipstick [127,128] or one dipstick measurement >/=2+) [129]. Severe preeclampsia was diagnosed according to the criteria proposed by the American College of Obstetricians and Gynecologists (ACOG) committee [128]. Patients with preeclampsia were sub-classified as either early-onset (<34 weeks) or late onset (>/=34 weeks) disease according to the gestational age at which preeclampsia was diagnosed. A neonate was defined as SGA when the birth weight was below the 10th percentile for gestational age according to the reference range proposed by Alexander et al. [130]. Patients were considered to have a normal pregnancy if they met the following criteria: (1) no medical, obstetrical, or surgical complications; (2) absence of labor at the time of venipuncture; and (3) delivery of a normal term (>/=37 weeks) infant whose birth weight was between the 10th and 90th percentile for gestational age [130]. The non-pregnant group consisted of healthy volunteers not taking oral contraceptives whose blood was drawn in the secretory phase of the menstrual cycle.
IP-10 (CXCL10) determinations
Specific and sensitive enzyme-linked immunoassays were used to determine concentrations of IP-10 in human maternal serum. Immunoassays for IP-10 were obtained from R&D Systems (Minneapolis, MN, USA). Briefly, maternal serum samples were incubated in duplicate wells of the microtiter plates, pre-coated with a monoclonal antibody specific for IP-10. During this incubation step, any IP-10 present in the standards or maternal serum is bound by the immobilized antibodies. After repeated washing and aspiration to remove all unbound substances, an enzyme-linked polyclonal antibody specific for IP-10 was added to the wells. Following a wash to remove excess and unbound materials, a substrate solution was added to the wells and color developed in proportion to the amount of IP- 10 bound in the initial step. The color development was stopped with the addition of an acid solution and the intensity of color was read using a programmable spectrophotometer (SpectraMax M2, Molecular Devices, Sunnyvale, CA, USA). The concentrations of IP-10 in serum samples were determined by interpolation from individual standard curves composed of recombinant human IP-10. The calculated inter- and intra-assay coefficients of variation for IP-10 immunoassays in our laboratory were 7.99% and 4.12%, respectively. The lower limit of detection (sensitivity) was calculated to be 5.01 pg/mL.
Statistical analysis
The Kolmogorov-Smirnov test was used to determine whether the data were normally distributed. The Spearman’s correlation test was used in assessing the relationship between maternal serum concentration of IP-10 and gestational age at blood draw in patients with normal pregnancies. Comparisons among groups were performed using Kruskal-Wallis tests with post-hoc analysis for continuous variables, and Chi-square or Fisher’s exact test for categorical variables. A p value < 0.05 was considered statistically significant. The statistical package used was SPSS v.12.0 (SPSS Inc., Chicago, IL, USA). Results
Three hundred sixteen patients were included in this study. The demographic and clinical characteristics of the study groups are displayed in Table I.
Among patients with preeclampsia, 63% (63/100) were classified as early-onset and 88% (88/100) as severe preeclampsia. In 75.6% (59/ 78) of patients who delivered an SGA neonate, the birth weight was below the 5th percentile. IP-10 was detectable in the serum of all subjects.
Table I. Clinical and obstetrical characteristics of the study groups.
Serum concentration of IP-10 in pregnancy
Patients with normal pregnancies had a significantly higher median serum concentration of IP-10 than non-pregnant women (median 116.1 pg/mL, range 40.7-1314.3 vs. median 90.3 pg/mL, range 49.2- 214.7, respectively; p = 0.002) (see Figure 1). There was no significant correlation between maternal serum IP-10 concentration and gestational age at blood draw (r = 0.039; p = 0.7).
IP-10 in preeclampsia
Patients with preeclampsia had a higher median serum concentration of IP-10 than normal pregnant women and those who delivered an SGA neonate (preeclampsia: median 156.4 pg/mL, range 47.4-645.9; normal pregnancy: median 116.1 pg/mL, range 40.7- 1314.3; SGA: median 122.4 pg/mL, range 37.3-693.5; p < 0.05 for both comparisons). In contrast, there were no significant differences in the maternal serum median IP-10 concentrations between patients who delivered an SGA neonate and those with normal pregnancies (SGA: median 122.4 pg/mL, range 37.3-693.5 vs. normal pregnancy: median 116.1 pg/mL, range 40.7-1314.3; p > 0.05) (see Figure 2).
Among patients with preeclampsia, no significant differences in serum concentrations of IP-10 were observed between patients with early onset and late onset disease (early onset preeclampsia: median 155.6 pg/mL, range 47.5-645.9 vs. late onset preeclampsia: median 165.2 pg/mL, range 64.7-401.1; p = 0.4). Similarly, there were no significant differences between mild and severe preeclampsia (mild preeclampsia: median 177.7 pg/mL, range 79.4-401.1 vs. severe preeclampsia: median 155.5 pg/mL, range 47.5-645.9; p = 0.56).
Figure 1. Serum concentrations of CXCL10/IP-10 in non-pregnant women and in patients with normal pregnancies. Patients with normal pregnancies had a significantly higher median serum concentration of IP-10 than non-pregnant women (median 116.1 pg/mL, range 40.7- 1314.3 vs. median 90.3 pg/mL, range 49.2-214.7, respectively; p = 0.002).
Among patients who delivered an SGA neonate, there were no significant differences in serum concentrations of IP-10 between patients who delivered a neonate with a birth weight at < 5th percentile and those who delivered a neonate with a birth weight between the 5th and 9th percentile (SGA <5th percentile: median 126.7 pg/mL, range 37.3-693.5 vs. SGA 5th-9th percentile: median 108.7 pg/mL, range 40.7-349.5; p = 0.2).
Using analysis of covariance (ANCOVA), only diagnostic groups but not storage time had a significant effect on the serum concentrations of CXCL10 (storage time: p = 0.5; diagnostic groups: p < 0.001). Storage time did not have a significant effect on the serum concentrations of CXCL10 even when gestational age at blood draw was included in the analysis (storage time: p = 0.72; diagnostic groups: p = 0.02; gestational age at blood draw: p = 0.5).
Discussion
Principal findings of this study
(1) The median serum concentration of IP-10 in normal pregnancy was higher than that of non-pregnant women; (2) there was no relationship between gestational age and the maternal serum concentration of IP-10; (3) preeclampsia, but not SGA, was associated with a higher median concentration of maternal serum IP- 10. These results are novel and suggest that IP-10 may participate in the pathophysiology of preeclampsia.
What is CXCL10/IP-10?
IP-10 (CXCL10) is a chemokine of the CXC family [48], which was first described as the product of a gene induced in response to recombinant IFN-gamma in several cell populations, including U937 histiocytic lymphoma, human fibroblasts, mononuclear and endothelial cells [61]. The principal biological activity of chemotactic cytokines, such as IP-10, is regulation and control of the basal homeostatic and inflammatory leukocyte movement [57].
In addition, IP-10 has potent anti-angiogenic properties [54,56,57,59], promotes adhesion, migration, and invasion of trophoblast cell [131,132], has an inhibitory effect on early hematopoietic progenitors [133], and regulates intestinal crypt cell renewal in both physiologic conditions and during mucosal regeneration following injury [134].
The biological properties of IP-10 are mediated through the interaction with a transmembrane G protein-coupled receptor, CXCR3 [135,136] shared by two other IFN-gamma inducible CXC chemokines- CXCL9 (MIG) and CXCL11 (I-TAQ-whose distinct biological activities are related to different transduction pathways [137-140].
Figure 2. Maternal serum concentrations of CXCL10/IP-10 among the study groups. Patients with preeclampsia had a significantly higher median serum concentration of IP-10 than normal pregnant women (median 156.4 pg/mL, range 47.4-645.9 vs. median 116.1 pg/mL, range 40.7-1314.3, respectively; p < 0.05) and than patients who delivered a small for gestational age (SGA) neonate (median 156.4 pg/mL, range 47.4-645.9 vs. median 122.4 pg/mL, range 37.3-693.5, respectively; p < 0.05). No significant differences were found in maternal serum median IP-10 concentrations between patients who delivered an SGA neonate and those with normal pregnancies (median 122.4 pg/ mL, range 37.3-693.5 vs. median 116.1 pg/mL, range 40.7-1314.3, respectively; p > 0.05).
Factors controlling the expression of IP-10 and cell sources
IP-10 gene and protein expression is modulated by pro- inflammatory stimuli. Indeed, IFN-gamma is an inducer of the gene, and protein expression of this chemokine by mononuclear cells [61], neutrophils [69,70], eosinophils [70,141], keratinocytes [61,64, 65,67], fibroblasts [61], endothelial cells [61-63], pancreatic beta cells [71,72], and animal astrocytes/ microglia [66,68], TNF-alpha [70,73-79], interleukin (IL)-1beta [70,71], as well as viral [66,81,82] and microbial products [70,80,83-85], can also stimulate the production of IL-10.
Activation of the nuclear factor-kappa B (NF-kB) pathway [81,82,86-89] through ligation of pattern recognition receptors (TLR4 [83] and TLR3 [89]) can also upregulate gene and protein expression of IP-10. Interestingly, IP-10 is considered an ‘NF-kB responsive gene’ [142].
IP-10 in inflammation
The pro-inflammatory activity of IP-10 includes: chemotaxis and endothelial adhesion of activated T cells [143] as well as chemotaxis [144] and enhancement of natural killer (NK) cell- mediated cytolysis [144]. However, this chemokine is a poor neutrophil activator [143,145] and there is controversy about its effects on both monocytes [135,143] and B cells [146,147].
CXCR3 receptor expression on T lymphocytes is selective for activated cells [135,143,146,148-151]. Interestingly, analysis of polarized T lymphocytes using specific monoclonal antibodies has demonstrated high CXCR3 expression (mRNA and protein) on Th1 cells and low on Th2 cells [149,152], and this receptor has, therefore, been proposed as a useful clinical marker of circulating Th1-type cells [149,153]. Further evidence that IP-10 promotes a Th1-like dominance is the observation that, following stimulation with phorbol-12-myristate 13-acetate (PMA) and ionomycin, the production of IFN-gamma (a Th1-type chemokine) resides exclusively in CXCR3- expressing memory CD4+ T cells, whereas the production of Th2-type cytokines is mainly observed in those CXCR3-negative [153]. In addition, in vitro assays show that, in polyclonally stimulated T cells, recombinant IP-10 selectively enhances IFN-gamma protein synthesis, while having no effect on IL-4 production [154]. The capability of IP-10 to enhance its own inducer, IFN-gamma supports the hypothesis that a positive amplification loop between IP-10 and IFN-gamma also exists in vivo [154].
IP-10 in pathologic states
IP-10 has been implicated in states characterized by a prominent T cell response [155,156], particularly when a Th1/Th2 imbalance is involved, including: (1) multiple sclerosis, where the serum and cerebrospinal fluid concentrations of IP10 correlate with the disease activity and CXCR3 expression is detectable on the majority of CNS-infiltrating lymphocytes [104-108]; (2) herpetic encephalitis in mice [157,158]; (3) experimental autoimmune encephalomyelitis [159-162]; (4) inflammatory bowel/colon disease [134,163,164]; (5) chronic hepatitis C, in which IP-10 serum concentrations are related to the inflammatory activity and the response to therapy [165]; (6) Sjogren’s syndrome, where the expression of IP-10 mRNA is significantly up-regulated in salivary glands (p < 0.01) and IP-10 has a potential role in the accumulation of T cells infiltrates [166]; (7) type 1 diabetes, whose immunopathogenesis is likely to be linked to the IP-10 [109,110] property of enhancing the traffic of auto-aggressive cells to the pancreas [167] and imprinting a pattern for the subsequent development of the autoimmune disease [109]; (8) Graves' disease [111-114] and autoimmune thyroiditis [115,116] where high concentrations of IP-10 have been detected in the serum of affected individuals; and (9) systemic lupus erythematosus, where IP- 10 plasma concentrations not only are higher than in non-diseased individuals but also correlate with the disease activity [168].
IP-10 as an inhibitor of angiogenesis
IP-10 has potent anti-angiogenic properties [54,56,57,59]. Several mechanisms have been proposed to mediate these activities: (1) interaction with the CXCR3 receptor [169,170]; (2) binding to a cell surface heparan sulfate site shared with platelet factor 4 [124]; (3) interference with the pro-angiogenic activity of basic fibroblast growth factor (bFGF) and IL-8 [124]; and (4) through another high affinity receptor (different from CXCR3 and glycosaminoglycans) [171]. The IP-10 receptor CXCR3 (mRNA and protein) has been localized on the endothelial cells of several human tissues (kidney, gut, liver, thyroid, and thymus) [170,172]. Interestingly, the endothelial cell expression is cell cycle- dependent [170]. Indeed, staining is remarkably more frequent in conditions of activation and in the presence of a high proliferative rate (such as in inflamed and neoplastic tissues rather than in normal tissues), particularly during the S/G2-M phase of the endothelial cell cycle [170]. Romagnani et al. [170] suggested that these results represent evidence that CXCR3 is mediating the angiostatic activity of IP-10, thus controlling endothelial cell proliferation. In addition, two distinct isoforms of this receptor, CXCR3-A and CXCR3-B (both products of alternative splicing) are likely to mediate opposite effects on cellular proliferation [173,174].
A specific region of the IP-10 molecule has been linked to its anti-angiogenic effect. Indeed, an aminoterminal truncated form of IP-10, resulting from a post-translational processing, has impaired receptor signaling and lymphocyte chemotaxis, but retains anti- angiogenic properties [175].
Evidence in support of the anti-angiogenic role of IP-10 includes: (1) IP-10, added to human umbilical cord vein endothelial cells (HUVECs), cultured on a matrigel substrate, inhibits their differentiation into tube-like structures in a dose-dependent fashion [125] and reduces the extent of the neo-vascular network [126]; (2) IP-10, both in vitro [123,124] and in vivo [123,125] inhibits in a dose-dependent manner the pro-angiogenic effects of IL- 8 and bFGF, including endothelial cell chemotaxis and proliferation as well as neovascularization in animal models of angiogenesis (rat cornea [123] or matrigel injected in the subcutaneous tissue of nude mice [125]); (3) recombinant IP-10 can inhibit [^sup 3^H]-thymidine incorporation into HUVECs cultured with bFGF [124]. Further support of the anti-angiogenic properties of IP-10 is the role attributed to this chemokine in the pathogenesis of pulmonary fibrosis. Indeed, the lower production of IP-10 in lung tissue of patients with pulmonary fibrosis, as determined by ELISA, has been proposed to contribute to the greater angiogenic activity observed in these patients [117,119]. Additionally, when IP-10 is administered systemically to bleomycin pre-treated mice, it inhibits fibroplasia and deposition of extracellular matrix through the regulation of the local angiogenesis, resulting in a significant attenuation of the severity of the pulmonary fibrosis induced by this chemical [118]. This experimental evidence indicates that IP-10 has an important anti-angiogenic effect.
Evidence that the anti-angiogenic properties of IP-10 may have therapeutic value is the observation that it inhibits tumor growth. Observations in support of this include: (1) Burkitt’s tumors implanted into athymic mice, once injected or transfected with IP- 10, show histological evidence of tissue necrosis, capillary damage, intimal thickening, and vascular thrombosis [176]; (2) high serum concentrations of IP-10 have been reported in patients with lymphoproliferative disorders [177]; (3) tumors derived from IP-10 transduced melanoma cells have a reduced in vivo growth compared to those originating from parental or null-transduced cells (p = 0.0002) and the growth inhibition is associated with a marked reduction in microvessel density [178]; (4) the angiostatic activity of IP-10 in human melanoma cell line appears to depend on binding to CXCR3 [169]; (5) tumors originating from human fibrosarcoma cell lines, secreting IP-10 upon vector transduction, and implanted into mice, have histological evidence of a significantly lower number of micro vessels than controls (p = 0.01) [126]; (6) recombinant IP-10 injection into non-small cell lung cancers grown in SCID (severe combined immunodeficiency) mice reduces their angiogenesis, growth, and incidence of spontaneous metastasis, whilst IP-10 neutralization results in enhanced tumor-derived angiogenic activity. Furthermore, plasma or tumor-associated IP-10 concentrations are inversely correlated to tumor growth. Extracts from these tumors decrease the angiogenic activity in the corneal micropocket assay [179]; and (7) IP-10 contributes to the anti-angiogenic and anti-tumor properties of IL-12 [180-183].
In summary, this evidence indicates that IP-10 plays an important anti-angiogenic role.
IP-10 concentration in serum during normal pregnancy
The observation that normal pregnant women have a significantly higher median serum concentration of IP-10 than non-pregnant women is novel. This observation is consistent with the findings that normal pregnancy is associated with systemic intravascular inflammation [35,184-187]. Additional evidence in support of this view is the observation that pregnancy is associated with higher (serum or plasma) concentrations of IL-12 [186] and TNF-alpha [188] than the non-pregnant state.
IP-10, inflammation, and preedampsia
The observation that preeclampsia is associated with a higher median maternal serum concentration of IP-10 than SGA and normal pregnancy is novel, as well as being consistent with the view that preeclampsia is characterized by an exaggerated intravascular pro- inflammatory state [18,33-37].
Furthermore, accumulating evidence indicates that preeclampsia is associated with a predominant Th1 immune response, including: (1) high maternal plasma or serum concentrations of IL-2 [38], TNF- alpha [40,41,189-191], IFN-gamma [192], and IL-12 [42,193]; (2) low maternal plasma or serum concentrations of IL-10 [194] and of IL-4 [192] (although this topic is subject to some controversy) [195]; and (3) upregulation of mRNA and protein expression of IL-1beta [196] and TNF-alpha [196,197] in the placenta. Thus, the high maternal plasma concentration of this chemokine in patients with preeclampsia may represent yet another feature of a pro- inflammatory state or intravascular inflammation or even contribute to the generation of such state. Some evidence also indicates that SGA may be associated with intravascular inflammation [47]. However, the results presented herein do not implicate IP-10 in such process.
IP-10, preeclampsia, and anti-angiogenesis
An imbalance between pro-angiogenic and anti-angiogenic factors is involved in the pathophysiology of preeclampsia [25-27,198-218]. The group of Maynard and Karumanchi has recently made a major set of observations that favors this hypothesis [219]. Evidence in support of this includes: (1) overexpression of sVEGFR-1 mRNA and protein in placenta of patients with preeclampsia [27,219]; (2) higher median plasma/serum concentration of sVEGFR-1 in preeclampsia at the time of diagnosis than in patients with normal pregnancies [27,219,220] and the plasma concentration correlates with the severity of the disease [28]; (3) preeclampsia is associated with decreased plasma/ serum concentrations of VEGF and PIGF [25,211,219,221]; (4) serum of pregnant women with preeclampsia has anti-angiogenic effects in the endothelial cell tube formation bioassay and these effects can be restored by the addition of VEGF and PlGF [219]; and (5) administration of sVEGFR-1 to pregnant animals can induce the clinical manifestation of preeclampsia, including hypertension and proteinuria [219]. Moreover, these animals develop the pathologic finding of glomerular endotheliosis but not pathognomonic of preeclampsia [222,223]; and (6) preeclampsia is associated with a higher maternal serum concentration of endoglin at the time of the diagnosis or before the recognition of the disease [31,32].
Given the evidence that IP-10 has anti-angiogenic properties and that this is a feature of preeclampsia, we propose that elevated IP- 10 maternal serum concentrations may contribute to generating an anti-angiogenic state along with sVEGFR-1 and endoglin.
IP-10, preeclampsia, and allograft rejection
Allograft rejection has also been proposed as a mechanism of disease in preeclampsia [224]. Of note, there is growing evidence that IP-10 is also involved in the process of graft rejection, including: (1) CXCR3 deficient (-/-) mice are resistant to both acute and chronic allograft rejection [91]; (2) grafts from IP-10 (- /-) donors are less likely to undergo graft injury [90]; (3) treatment with anti-CXCR3 [91] or anti-CXCL10 [96] monoclonal antibodies results in prolongation of cardiac and small bowel allograft survival in mice; (4) clinical rejection of human renal [92,100], lung [93], cardiac [94,95], small bowel [99], and arteries [102] transplants is associated with intra-graft over expression of IP-10 and/or with intra-graft recruitment of CXCR3 positive T cells; and (5) pre-transplant serum [97,103] and urine [98,101] CXCL10 cellular mRNA or protein concentrations identify patients at risk for the development of acute rejection and/or chronic allograft nephropathy. Collectively, this evidence indicates that IP-10 is involved in the pathogenesis of allograft rejection. Whether or not this mechanism of disease operates in preeclampsia remains an interesting concept. Our observation that IP-10 is elevated in preeclampsia provides a possible link between preeclampsia and this potential pathologic process.
IP-10, preeclampsia, and atherosclerosis
Striking parallels exist between preeclampsia and atherosclerosis (see bibliography for details). Patients who develop preterm preeclampsia are at increased risk of death from coronary artery disease later in life [225]. Moreover, atherosis of the spiral arteries, a lesion observed in patients with preeclampsia, has striking similarities with that of coronary artery disease [226- 228]. The current view of atherosclerosis is that it is an inflammatory mediated process [229-231]. Th1 biased responses are thought to be pro-atherogenic, while Th2 biased immunoresponses confer atheroprotection [232]. Chemokine-dependent migration of cells through the inflamed endothelium into the arterial intima has been proposed to be critical in atherosclerosis [233-237]. There is evidence of a specific involvement of IP-10 in the pathogenesis of atherosclerosis and cardiovascular diseases. Such evidence includes: (1) patients with stable angiographically confirmed coronary heart disease have a higher mean IP-10 serum concentration than control patients, and the magnitude of the elevation correlates with the concentration of several acute-phase proteins (C-reactive proteins) or cytokines known to be central in the pathogenesis of atherosclerosis [122]; (2) serum IP-10 baseline concentrations are significantly higher in individuals who develop coronary heart disease (CHD) than in those who do not (follow-up of 11 years). After adjustment for cardiovascular and immunological risk factors, however, the observed relationship with IP-10 disappeared, suggesting that the association may be explained by other markers of inflammation and not be specific to IP-10. Nonetheless, this observation is important because it suggests that an elevation of IP- 10 precedes the development of CHD, thus strengthening the case that inflammation has a causal role rather being a consequence of atherosclerosis [121]; (3) CXCR3-bearing T cells and chemokines IP- 10, I-TAC, and MIG (all three IFN-gamma-induced) are present in atherosclerotic plaques [120].
Conclusions
Preeclampsia is associated with higher maternal plasma concentrations of IP-10 than normal pregnancy and SGA. These results suggest that this chemokine may contribute to both the exaggerated systemic inflammation and the anti-angiogenic state that characterize preeclampsia.
Acknowledgement
This research was supported by the Intramural Research Program of the National Institute of Child Health and Human Development, NIH, DHHS.
References
1. Brosens IA, Robertson WB, Dixon HG. The role of the spiral arteries in the pathogenesis of pre-eclampsia. J Pathol 1970;101:vi.
2. Brosens I, Dixon HG, Robertson WB. Fetal growth retardation and the arteries of the placental bed. Br J Obstet Gynaecol 1977;84:656-663.
3. De Wolf F, Brosens I, Renaer M. Fetal growth retardation and the maternal arterial supply of the human placenta in the absence of sustained hypertension. Br J Obstet Gynaecol 1980;87:678-685.
4. Khong TY, De Wolf F, Robertson WB, Brosens I. Inadequate maternal vascular response to placentation in pregnancies complicated by pre-eclampsia and by small-for-gestational age infants. Br J Obstet Gynaecol 1986;93:1049-1059.
5. Pijnenborg R, Anthony J, Davey DA, Rees A, Tiltman A, Vercruysse L, van Assche A. Placental bed spiral arteries in the hypertensive disorders of pregnancy. Br J Obstet Gynaecol 1991;98:648-655.
6. Meekins JW, Pijnenborg R, Hanssens M, McFadyen IR, van Assche A. A study of placental bed spiral arteries and trophoblast invasion in normal and severe pre-eclamptic pregnancies. Br J Obstet Gynaecol 1994;101:669-674.
7. Brosens JJ, Pijnenborg R, Brosens IA. The myometrial junctional zone spiral arteries in normal and abnormal pregnancies: A review of the literature. Am J Obstet Gynecol 2002;187:1416-1423.
8. Espinoza J, Romero R, Kim YM, Kusanovic JP, Hassan S, Erez O, Gotsch F, Than NG, Papp Z, Kim CJ. Normal and abnormal transformation of the spiral arteries during pregnancy. J Perinat Med 2006;34:447-458.
9. Campbell S, Diaz-Recasens J, Griffin DR, Cohen-Overbeek TE, Pearce JM, Willson K, Teague MJ. New Doppler technique for assessing uteroplacental blood flow. Lancet 1983;1:675-677.
10. Harrington KF, Campbell S, Bewley S, Bower S. Doppler velocimetry studies of the uterine artery in the early prediction of pre-eclampsia and intra-uterine growth retardation. Eur J Obstet Gynecol Reprod Biol 1991;42(Suppl): S14-S20.
11. Bower S, Schuchter K, Campbell S. Doppler ultrasound screening as part of routine antenatal scanning: Prediction of pre- eclampsia and intrauterine growth retardation. Br J Obstet Gynaecol 1993;100:989-994.
12. Harrington K, Cooper D, Lees C, Hecher K, Campbell S. Doppler ultrasound of the uterine arteries: The importance of bilateral notching in the prediction of pre-eclampsia, placental abruption or delivery of a small-for-gestational-age baby. Ultrasound Obstet Gynecol 1996;7:182-188.
13. Dekker GA, Sibai BM. Etiology and pathogenesis of preeclampsia: Current concepts. Am J Obstet Gynecol 1998; 179:1359- 1375.
14. Albaiges G, Missfelder-Lobos H, Lees C, Parra M, Nicolaides KH. One-stage screening for pregnancy complications by color Doppler assessment of the uterine arteries at 23 weeks’ gestation. Obstet Gynecol 2000;96: 559-564.
15. Papageorghiou AT, Yu CK, Bindra R, Pandis G, Nicolaides KH. Multicenter screening for pre-eclampsia and fetal growth restriction by transvaginal uterine artery Doppler at 23 weeks of gestation. Ultrasound Obstet Gynecol 2001;18:441-449.
16. Papageorghiou AT, Yu CK, Nicolaides KH. The role of uterine artery Doppler in predicting adverse pregnancy outcome. Best Pract Res Clin Obstet Gynaecol 2004; 18: 383-396.
17. Roberts JM, Taylor RN, Musci TJ, Rodgers GM, Hubel CA, McLaughlin MK. Preeclampsia: An endothelial cell disorder. Am J Obstet Gynecol 1989;161:1200-1204.
18. Redman CW, Sacks GP, Sargent IL. Preeclampsia: An excessive maternal inflammatory response to pregnancy. Am J Obstet Gynecol 1999;180:499-506.
19. Bretelle F, Sabatier F, Blann A, D’Ercole C, Boutiere B, Mutin M, Boubli L, Sampol J, Dignat-George F. Maternal endothelial soluble cell adhesion molecules with isolated small for gestational age fetuses: Comparison with preeclampsia. BJOG 2001;108:1277-1282.
20. Poston L, Chappell LC. Is oxidative stress involved in the aetiology of pre-eclampsia? Acta Paediatr Suppl 2001;90:3-5.
21. Johnson MR, Anim-Nyame N, Johnson P, Sooranna SR, Steer PJ. Does endothelial cell activation occur with intrauterine growth restriction? BJOG 2002;109:836-839.
22. Roberts JM, Lain KY. Recent insights into the pathogenesis of pre-eclampsia. Placenta 2002;23:359-372.
23. Redman CW, Sargent IL. Latest advances in understanding preeclampsia. Science 2005;308:1592-1594.
24. Ness RB, Sibai BM. Shared and disparate components of the pathophysiologies of fetal growth restriction and preeclampsia. Am J Obstet Gynecol 2006; 195:40-49.
25. Torry DS, Wang HS, Wang TH, Caudle MR, Tony RJ. Preeclampsia is associated with reduced serum levels of placenta growth factor. Am J Obstet Gynecol 1998; 179: 1539-1544.
26. Taylor RN, Grimwood J, Taylor RS, McMaster MT, Fisher SJ, North RA. Longitudinal serum concentrations of placental growth factor: Evidence for abnormal placental angiogenesis in pathologic pregnancies. Am J Obstet Gynecol 2003;188:177-182.
27. Tsatsaris V, Coffin F, Munaut C, Enchant JF, Pignon MR, Noel A, Schaaps JP, Cabrol D, Frankenne F, Foidart JM. Overexpression of the soluble vascular endothelial growth factor receptor in preeclamptic patients: Pathophysiological consequences. J Clin Endocrinol Metab 2003;88:5555-5563.
28. Chaiworapongsa T, Romero R, Espinoza J, Bujold E, Mee KY, Goncalves LF, Gomez R, Edwin S. Evidence supporting a role for blockade of the vascular endothelial growth factor system in the pathophysiology of preeclampsia. Young Investigator Award. Am J Obstet Gynecol 2004; 190:1541-1547.
29. Levine RJ, Maynard SE, Qian C, Lim KH, England LJ, Yu KF, Schisterman EF, Thadhani R, Sachs BP, Epstein FH, et al. Circulating angiogenic factors and the risk of preeclampsia. N Engl J Med 2004;350: 672-683.
30. Shibata E, Rajakumar A, Powers RW, Larkin RW, Gilmour C, Bodnar LM, Crombleholme WR, Ness RB, Roberts JM, Hubel CA. Soluble fms-like tyrosine kinasc 1 is increased in preeclampsia but not in normotensive pregnancies with small-for-gestational-age neonates: Relationship to circulating placental growth factor. J Clin Endocrinol Metab 2005;90:4895-4903.
31. Levine RJ, Lam C, Qian C, Yu KF, Maynard SE, Sachs BP, Sibai BM, Epstein FH, Romero R, Thadhani R, et al. Soluble endoglin and other circulating antiangiogenic factors in preeclampsia. N Engl J Med 2006;355: 992-1005.
32. Venkatesha S, Toporsian M, Lam C, Hanai J, Mammoto T, Kirn YM, Bdolah Y, Lim KH, Yuan HT, Libermann TA, et al. Soluble endoglin contributes to the pathogenesis of preeclampsia. Nat Med 2006; 12:642-649.
33. Barden A, Graham D, Beilin LJ, Ritchie J, Baker R, Walters BN, Michael CA. Neutrophil CD11B expression and neutrophil activation in pre-eclampsia. Clin Sci (Lond) 1997;92:37-44.
34. Haller H, Ziegler EM, Homuth V, Drab M, Eichhorn J, Nagy Z, Busjahn A, Vetter K, Luft FC. Endothelial adhesion molecules and leukocyte integrins in preeclamptic patients. Hypertension 1997;29:291-296.
35. Sacks GP, Studena K, Sargent K, Redman CW. Normal pregnancy and preeclampsia both produce inflammatory changes in peripheral blood leukocytes akin to those of sepsis. Am J Obstet Gynecol 1998;179:80-86.
36. Gervasi MT, Chaiworapongsa T, Pacora P, Naccasha N, Yoon BH, Maymon E, Romero R. Phenotypic and metabolic characteristics of monocytes and granulocytes in preeclampsia. Am J Obstet Gynecol 2001;185:792-797.
37. Chaiworapongsa T, Gervasi MT, Refuerzo J, Espinoza J, Yoshimatsu J, Berman S, Romero R. Maternal lymphocyte subpopulations (CD45RA+ and CD45RO+) in preeclampsia. Am J Obstet Gynecol 2002;187:889-893.
38. Sunder-Plassmann G, Derfler K, Wagner L, Stockenhuber F, Endler M, Nowotny C, Balcke P. Increased serum activity of interleukin-2 in patients with pre-eclampsia. J Autoimmun 1989;2:203- 205.
39. Greer IA, Lyall F, Perera T, Boswell F, Macara LM. Increased concentrations of cytokines interleukin-6 and interleukin-1 receptor antagonist in plasma of women with preeclampsia: A mechanism for endothelial dysfunction? Obstet Gynecol 1994;84:937-940. 40. Vince GS, Starkey PM, Austgulen R, Kwiatkowski D, Redman CW. Interleukin- 6, tumour necrosis factor and soluble tumour necrosis factor receptors in women with pre-eclampsia. Br J Obstet Gynaecol 1995;102: 20-25.
41. Conrad KP, Miles TM, Benyo DF. Circulating levels of immunoreactive cytokines in women with preeclampsia. Am J Reprod Immunol 1998;40:102-111.
42. Daniel Y, Kupferminc MJ, Baram A, Jaffa AJ, Fait G, Wolman I, Lessing JB. Plasma interleukin-12 is elevated in patients with preeclampsia. Am J Reprod Immunol 1998; 39:376-380.
43. Saito S, Umekage H, Sakamoto Y, Sakai M, Tanebe K, Sasaki Y, Morikawa H. Increased T-helper-1 -type immunity and decreased T- helper-2-type immunity in patients with preeclampsia. Am J Reprod Immunol 1999;41:297-306.
44. Saito S, Sakai M, Sasaki Y, Tanebe K, Tsuda H, Michimata T. Quantitative analysis of peripheral blood Th0, Th1, Th2 and the Th1:Th2 cell ratio during normal human pregnancy and preeclampsia. Clin Exp Immunol 1999;117:550-555.
45. Darmochwal-Kolarz D, Leszczynska-Gorzelak B, Rolinski J, Oleszczuk J. T helper 1- and T helper 2-type cytokine imbalance in pregnant women with pre-eclampsia. Eur J Obstet Gynecol Reprod Biol 1999;86:165-170.
46. Sakai M, Ogawa K, Shiozaki A, Yoneda S, Sasaki Y, Nagata K, Saito S. Serum granulysin is a marker for Th1 type immunity in pre- eclampsia. Clin Exp Immunol 2004; 136:114-119.
47. Bartha JL, Romero-Carmona R, Comino-Delgado R. Inflammatory cytokines in intrauterine growth retardation. Acta Obstet Gynecol Scand 2003;82:1099-1102.
48. Neville LF, Mathiak G, Bagasra O. The immunobiology of interferon-gamma inducible protein 10 kD (IP-10): A novel, pleiotropic member of the C-X-C chemokine superfamily. Cytokine Growth Factor Rev 1997;8:207-219.
49. Stricter RM, Polverini PJ, Kunkel SL, Arenberg DA, Burdick MD, Kasper J, Dzuiba J, Van Damme J, WaIz A, Marriott D, et al. The functional role of the ELR motif in CXC chemokine-mediated angiogenesis. J Biol Chem 1995; 270:27348-27357.
50. Stricter RM, Polverini PJ, Arenberg DA, Kunkel SL. The role of CXC chemokines as regulators of angiogenesis. Shock 1995;4:155- 160.
51. Stricter RM, Polverini PJ, Arenberg DA, Walz A, Opdenakker G, Van Damme J, Kunkel SL. Role of C-X-C chemokines as regulators of angiogenesis in lung cancer. J Leukoc Biol 1995;57:752-762.
52. Rollins BJ. Chemokines. Blood 1997;90:909-928.
53. Luster AD. Chemokines-chemotactic cytokines that mediate inflammation. N Engl J Med 1998;338:436-445.
54. Belperio JA, Keane MP, Arenberg DA, Addison CL, Ehlert JE, Burdick MD, Stricter RM. CXC chemokines in angiogenesis. J Leukoc Biol 2000;68:1-8.
55. Rossi D, Zlotnik A. The biology of chemokines and their receptors. Annu Rev Immunol 2000;18:217-242.
56. Bernardini G, Ribatti D, Spinetti G, Morbidelli L, Ziche M, Santoni A, Capogrossi MC, Napolitano M. Analysis of the role of chemokines in angiogenesis. J Immunol Methods 2003;273:83-101.
57. Rosenkilde MM, Schwartz TW. The chemokine system-a major regulator of angiogenesis in health and disease. APMIS 2004;112:481- 495.
58. Romagnani P, Lasagni L, Annunziato F, Serio M, Romagnani S. CXC chemokines: The regulatory link between inflammation and angiogenesis. Trends Immunol 2004;25:201-209.
59. Stricter RM, Burdick MD, Gomperts BN, Belperio JA, Keane MP. CXC chemokines in angiogenesis. Cytokine Growth Factor Rev 2005;16:593-609.
60. Charo IF, Ransohoff RM. The many roles of chemokines and chemokine receptors in inflammation. N Engl J Med 2006;354:610-621.
61. Luster AD, Unkeless JC, Ravetch JV. Gamma-interferon transcriptionally regulates an early-response gene containing homology to platelet proteins. Nature 1985;315: 672-676.
62. Luster AD, Ravetch JV. Biochemical characterization of a gamma interferon-inducible cytokine (IP-10). J Exp Med 1987;166:1084- 1097.
63. Luster AD, Ravetch JV. Genomic characterization of a gamma- interferon-inducible gene (IP-10) and identification of an interferon-inducible hypersensitive site. Mol Cell Biol 1987;7:3723- 3731.
64. Kaplan G, Luster AD, Hancock G, Cohn ZA. The expression of a gamma interferon-induced protein (IP-10) in delayed immune responses in human skin. J Exp Med 1987; 166:1098-1108.
65. Gottlieb AB, Luster AD, Posnett DN, Carter DM. Detection of a gamma interferon-induced protein IP-10 in psoriatic plaques. J Exp Med 1988;168:941-948.
66. Vanguri P, Farber JM. IFN and virus-inducible expression of an immediate early gene, crg-2/IP-10, and a delayed gene, I-A alpha in astrocytes and microglia. J Immunol 1994; 152: 1411-1418.
67. Sarris AH, Esgleyes-Ribot T, Crow M, Broxmeyer HE, Karasawas N, Pugh W, Grossman D, Deisseroth A, Duvic M. Cytokine loops involving interferon-gamma and IP-10, a cytokine chemotactic for CD4+ lymphocytes: An explanation for the epidermotropism of cutaneous T-cell lymphoma? Blood 1995;86:651-658.
68. Vanguri P. Interferon-gamma-inducible genes in primary glial cells of the central nervous system: Comparisons of astrocytes with microglia and Lewis with brown Norway rats. J Neuroimmunol 1995;56:35-43.
69. Cassatella MA, Gasperini S, Calzetti F, Bertagnin A, Luster AD, McDonald PP. Regulated production of the interferon-gamma- inducible protein-10 (IP-10) chemokine by human neutrophils. Eur J Immunol 1997;27: 111-115.
70. Gasperini S, Marchi M, Calzetti F, Laudanna C, Vicentini L, Olsen H, Murphy M, Liao F, Farber J, Cassatella MA. Gene expression and production of the monokine induced by IFN-gamma (MIG), IFN- inducible T cell alpha chemoattractant (I-TAC), and IFN-gamma- inducible protein-10 (IP-10) chemokines by human neutrophils. J Immunol 1999; 162: 4928-4937.
71. Frigerio S, Junt T, Lu B, Gerard C, Zumsteg U, Hollander GA, Piali L. Beta cells are responsible for CXCR3-mediated T-cell infiltration in insulins. Nat Med 2002;8:1414-1420.
72. Cardozo AK, Proost P, Gysemans C, Chen MC, Mathieu C, Eizirik DL. IL-1beta and IFN-gamma induce the expression of diverse chemokines and IL-15 in human and rat pancreatic islet cells, and in islets from pre-diabetic NOD mice. Diabetologia 2003;46:255-266.
73. Narumi S, Yoneyama H, Inadera H, Nishioji K, Itoh Y, Okanoue T, Matsushima K. TNF-alpha is a potent inducer for IFN-inducible protein-10 in hepatocytes and unaffected by GM-CSF in vivo, in contrast to IL-1beta and IFN-gamma. Cytokine 2000; 12: 1007-1016.
74. Kraft M, Riedel S, Maaser C, Kucharzik T, Steinbuechel A, Domschke W, Luegering N. IFN-gamma synergizes with TNF-alpha but not with viable H. pylon in up-regulating CXC chemokine secretion in gastric epithelial cells. Clin Exp Immunol 2001;126:474-481.
75. Algood HM, Lin PL, Yankura D, Jones A, Chan J, Flynn JL. TNF influences chemokine expression of macrophages in vitro and that of CD11b+ cells in vivo during Mycobacterium tuberculosis infection. J Immunol 2004; 172: 6846-6857.
76. Villagomez MT, Bae SJ, Ogawa I, Takenaka M, Katayama I. Tumour necrosis factor-alpha but not interferon-gamma is the main inducer of inducible protein-10 in skin fibroblasts from patients with atopic dermatitis. Br J Dermatol 2004; 150:910-916.
77. Hardaker EL, Bacon AM, Carlson K, Roshak AK, Foley JJ, Schmidt DB, Buckley PT, Comegys M, Panettieri RA Jr, Sarau HM, et al. Regulation of TNF-alpha- and IFN-gamma-induced CXCL10 expression: Participation of the airway smooth muscle in the pulmonary inflammatory response in chronic obstructive pulmonary disease. FASEB J 2004; 18: 191-193.
78. Sheng WS, Hu S, Ni HT, Rowen TN, Lokensgard JR, Peterson PK. TNF-alpha-induced chemokine production and apoptosis in human neural precursor cells. J Leukoc Biol 2005;78:1233-1241.
79. Berthier-Vergnes O, Bermond F, Flacher V, Massacrier C, Schmitt D, Peguet-Navarro J. TNF-alpha enhances phenotypic and functional maturation of human epidermal Langerhans cells and induces IL-12 p40 and IP-10/CXCL-10 production. FEBS Lett 2005;579:3660-3668.
80. Shin HS, Drysdale BE, Shin ML, Noble PW, Fisher SN, Paznekas WA. Definition of a lipopolysaccharide-responsive element in the 5′- flanking regions of MuRantes and crg-2. Mol Cell Biol 1994;14:2914- 2925.
81. Nazar AS, Cheng G, Shin HS, Brothers PN, Dhib-Jalbut S, Shin ML, Vanguri P. Induction of IP-10 chemokine promoter by measles virus: Comparison with interferon-gamma shows the use of the same response element but with differential DNA-protein binding profiles. J Neuroimmunol 1997;77:116-127.
82. Cheng G, Nazar AS, Shin HS, Vanguri P, Shin ML. IP-10 gene transcription by virus in astrocytes requires cooperation of ISRE with adjacent kappaB site but not IRF-1 or viral transcription. J Interferon Cytokine Res 1998; 18: 987-997.
83. Gasper NA, Petty CC, Schrum LW, Marriott I, Host KL. Bacterium-induced CXCL10 secretion by osteoblasts can be mediated in part through toll-like receptor 4. Infect Immun 2002;70:4075-4082.
84. Shen Q, Zhang R, Bhat NR. MAP kinase regulation of IP10/ CXCL10 chemokine gene expression in microglial cells. Brain Res 2006;1086:9-16.
85. Durand SH, Flacher V, Romeas A, Carrouel F, Colomb E, Vincent C, Magloire H, Couble ML, Bleicher F, Staquet MJ, et al. Lipoteichoic acid increases TLR and functional chemokine expression while reducing dentin formation in in vitro differentiated human odontoblasts. J Immunol 2006; 176: 2880-2887.
86. Ohmori Y, Hamilton TA. Cooperative interaction between interferon (IFN) stimulus response element and kappa B sequence motifs controls IFN gamma- and lipopolysaccharide-stimulated transcription from the murine IP-10 promoter. J Biol Chem 1993;268:6677-6688.
87. Ohmori Y, Hamilton TA. The interferon-stimulated response element and a kappa B site mediate synergistic induction of murine IP-10 gene transcription by IFN-gamma and TNF-alpha. J Immunol 1995;154:5235-5244.
88. Osawa Y, Iho S, Takauji R, Takatsuka H, Yamamoto S, Takahashi T, Horiguchi S, Urasaki Y, Matsuki T, Fujieda S. Collaborative action of NF-kappaB and p38 MAPK is involved in CpG DNA-induced IFN- alpha and chemokine production in human plasmacytoid dendritic cells. J Immunol 2006;177:4841-4852. 89. Park C, Lee S, Cho IH, Lee HK, Kirn D, Choi SY, Oh SB, Park K, Kirn JS, Lee SJ. TLR3-mediated signal induces proinflammatory cytokine and chemokine gene expression in astrocytes: Differential signaling mechanisms of TLR3- induced IP-10 and IL-8 gene expression. Glia 2006;53: 248-256.
90. Hancock WW, Gao W, Csizmadia V, Faia KL, Shemmeri N, Luster AD. Donor-derived IP-10 initiates development of acute allograft rejection. J Exp Med 2001;193:975-980.
91. Hancock WW, Lu B, Gao W, Csizmadia V, Faia K, King JA, Smiley ST, Ling M, Gerard NP, Gerard C. Requirement of the chemokine receptor CXCR3 for acute allograft rejection. J Exp Med 2000;192:1515-1520.
92. Segerer S, Cui Y, Eitner F, Goodpaster T, Hudkins KL, Mack M, Cartron JP, Colin Y, Schlondorff D, Alpers CE. Expression of chemokines and chemokine receptors during human renal transplant rejection. Am J Kidney Dis 2001; 37:518-531.
93. Agostini C, Calabrese F, Rea F, Facco M, Tosoni A, Loy M, Binotto G, Valente M, Trentin L, Semenzato G. Cxcr3 and its ligand CXCL10 are expressed by inflammatory cells infiltrating lung allografts and mediate chemotaxis of T cells at sites of rejection. Am J Pathol 2001;158: 1703-1711.
94. Melter M, Exeni A, Reinders ME, Fang JC, McMahon G, Ganz P, Hancock WW, Briscoe DM. Expression of the chemokine receptor CXCR3 and its ligand IP-10 during human cardiac allograft rejection. Circulation 2001;104: 2558-2564.
95. Zhao DX, Hu Y, Miller GG, Luster AD, Mitchell RN, Libby P. Differential expression of the IFN-gamma-inducible CXCR3-binding chemokines, IFN-inducible protein 10, monokine induced by IFN, and IFN-inducible T cell alpha chemoattractant in human cardiac allografts: Association with cardiac allograft vasculopathy and acute rejection. J Immunol 2002;169:1556-1560.
96. Zhang Z, Kaptanoglu L, Haddad W, Ivancic D, Alnadjim Z, Hurst S, Tishler D, Luster AD, Barren TA, Fryer J. Donor T cell activation initiates small bowel allograft rejection through an IFN-gamma- inducible protein-10-dependent mechanism. J Immunol 2002;168: 3205- 3212.
97. Rotondi M, Rosati A, Buonamano A, Lasagni L, Lazzeri E, Pradella F, Fossombroni V, Cirami C, Liotta F, La Villa G, et al. High pretransplant serum levels of CXCL10/IP-10 are related to increased risk of renal allograft failure. Am J Transplant 2004;4:1466-1474.
98. Hu H, Aizenstein BD, Puchalski A, Burmania JA, Hamawy MM, Knechtle SJ. Elevation of CXCR3-binding chemokines in urine indicates acute renal-allograft dysfunction. Am J Transplant 2004;4:432-437.
99. Zhang Z, Kaptanoglu L, Tang Y, Ivancic D, Rao SM, Luster A, Barren TA, Fryer J. IP-10-induced recruitment of CXCR3 host T cells is required for small bowel allograft rejection. Gastroenterology 2004; 126:809-818.
100. Panzer U, Reinking RR, Steinmetz OM, Zahner G, Sudbeck U, Fehr S, Pfalzer B, Schneider A, Thaiss F, Mack M, et al. CXCR3 and CCR5 positive T-cell recruitment in acute human renal allograft rejection. Transplantation 2004;78:1341-1350.
101. Tatapudi RR, Muthukumar T, Dadhania D, Ding R, Li B, Sharma VK, Lozada-Pastorio E, Seetharamu N, Hartono C, Serur D, et al. Noninvasive detection of renal allograft inflammation by measurements of mRNA for IP-10 and CXCR3 in urine. Kidney Int 2004;65: 2390-2397.
102. Burns WR, Wang Y, Tang PC, Ranjbaran H, lakimov A, Kirn J, Cuffy M, Bai Y, Pober JS, Tellides G. Recruitment of CXCR3+ and CCR5+ T cells and production of interferon-gamma-inducible chemokines in rejecting human arteries. Am J Transplant 2005;5:1226- 1236.
103. Lazzeri E, Rotondi M, Mazzinghi B, Lasagni L, Buonamano A, Rosati A, Pradella F, Fossombroni V, La Villa G, Gacci M, et al. High CXCL10 expression in rejected kidneys and predictive role of pretransplant serum CXCL10 for acute rejection and chronic allograft nephropathy. Transplantation 2005;79:1215-1220.
104. Sorensen TL, Tani M, Jensen J, Pierce V, Lucchinetti C, Folcik VA, Qin S, Rottman J, Sellebjerg F, Stricter RM, et al. Expression of specific chemokines and chemokine receptors in the central nervous system of multiple sclerosis patients. J Clin Invest 1999;103:807-815.
105. Balashov KE, Rottman JB, Weiner HL, Hancock WW. CCR5(+) and CXCR3(+) T cells are increased in multiple sclerosis and their ligands MIP-1alpha and IP-10 are expressed in demyelinating brain lesions. Proc Natl Acad Sci U S A 1999;96:6873-6878.
106. Simpson JE, Newcombe J, Cuzner ML, Woodroofe MN. Expression of the interferon-gamma-inducible chemokines IP-10 and Mig and their receptor, CXCR3, in multiple sclerosis lesions. Neuropathol Appl Neurobiol 2000;26: 133-142.
107. Franciotta D, Martino G, Zardini E, Furlan R, Bergamaschi R, Andreoni L, Cosi V. Serum and CSF levels of MCP-1 and IP-10 in multiple sclerosis patients with acute and stable disease and undergoing immunomodulatory therapies. J Neuroimmunol 2001;115:192- 198.
108. Trebst C, Ransohoff RM. Investigating chemokines and chemokine receptors in patients with multiple sclerosis: Opportunities and challenges. Arch Neurol 2001;58: 1975-1980.
109. Christen U, McGavern DB, Luster AD, Von Herrath MG, Oldstone MB. Among CXCR3 chemokines, IFN-gamma-inducible protein of 10 kDa (CXC chemokine ligand (CXCL) 10) but not monokine induced by IFN- gamma (CXCL9) imprints a pattern for the subsequent development of autoimmune disease. J Immunol 2003;171: 6838-6845.
110. Christen U, Von Herrath MG. IP-10 and type 1 diabetes: A question of time and location. Autoimmuniry 2004;37: 273-282.
111. Romagnani P, Rotondi M, Lazzeri E, Lasagni L, Francalanci M, Buonamano A, Milani S, Vitti P, Chiovato L, Tonacchera M, et al. Expression of IP-10/ CXCLlO and MIG/CXCL9 in the thyroid and increased levels of IP-10/CXCL10 in the serum of patients with recent-onset Graves’ disease. Am J Pathol 2002;161:195-206.
112. Antonelli A, Fallahi P, Rotondi M, Ferrari SM, Serio M, Miccoli P. Serum levels of the interferon-gamma-inducible alpha chemokine CXCL10 in patients with active Graves’ disease, and modulation by methimazole therapy and thyroidectomy. Br J Surg 2006;93:1226-1231.
113. Antonelli A, Fallahi P, Rotondi M, Ferrari SM, Romagnani P, Grosso M, Ferrannini E, Serio M. Increased serum CXCL10 in Graves’ disease or autoimmune thyroiditis is not associated with hyper- or hypothyroidism per se, but is specifically sustained by the autoimmune, inflammatory process. Eur J Endocrinol 2006;154:651- 658.
114. Antonelli A, Rotondi M, Fallahi P, Romagnani P, Ferrari SM, Barani L, Ferrannini E, Serio M. Increase of interferon-gamma- inducible CXC chemokine CXCL10 serum levels in patients with active Graves’ disease, and modulation by methimazole therapy. Clin Endocrinol (Oxf) 2006;64:189-195.
115. Antonelli A, Rotondi M, Fallahi P, Romagnani P, Ferrari SM, Buonamano A, Ferrannini E, Serio M. High levels of circulating CXC chemokine ligand 10 are associated with chronic autoimmune thyroiditis and hypothyroidism. J Clin Endocrinol Metab 2004;89:5496- 5499.
116. Antonelli A, Rotondi M, Fallahi P, Romagnani P, Ferrari SM, Paolicchi A, Ferrannini E, Serio M. Increase of interferon-gamma inducible alpha chemokine CXCL10 but not beta chemokine CCL2 serum levels in chronic autoimmune thyroiditis. Eur J Endocrinol 2005; 152: 171-177.
117. Keane MP, Arenberg DA, Lynch JP III, Whyte RI, Iannettoni MD, Burdick MD, Wilke CA, Morris SB, Glass MC, DiGiovine B, et al. The CXC chemokines, IL-8 and IP-10, regulate angiogenic activity in idiopathic pulmonary fibrosis. J Immunol 1997; 159: 1437-1443.
118. Keane MP, Belperio JA, Arenberg DA, Burdick MD, Xu ZJ, Xue YY, Stricter RM. IFN-gamma-inducible protein-10 attenuates bleomycin- induced pulmonary fibrosis via inhibition of angiogenesis. J Immunol 1999; 163: 5686-5692.
119. Stricter RM, Belperio JA, Keane MP. CXC chemokines in angiogenesis related to pulmonary fibrosis. Chest 2002;122: 298S- 301S.
120. Mach F, Saury A, Iarossi AS, Sukhova GK, Neote K, Ubby P, Luster AD. Differential expression of three T lymphocyte-activating CXC chemokines by human atheroma-associated cells. J Clin Invest 1999;104: 1041-1050.
121. Herder C, Baumert J, Thorand B, Martin S, Lowel H, Kolb H, Koenig W. Chemokines and incident coronary heart disease: Results from the MONICA/KORA Augsburg case-cohort study, 1984-2002. Arterioscler Thromb Vasc Biol 2006;26:2147-2152.
122. Rothenbacher D, Muller-Scholze S, Herder C, Koenig W, Kolb H. Differential expression of chemokines, risk of stable coronary heart disease, and correlation with established cardiovascular risk markers. Arterioscler Thromb Vasc Biol 2006;26:194-199.
123. Stricter RM, Kunkel SL, Arenberg DA, Burdick MD, Polverini PJ. Interferon gamma-inducible protein 10 (IP-10), a member of the C- X-C chemokine family, is an inhibitor of angiogenesis. Biochem Biophys Res Commun 1995;210:51-57.
124. Luster AD, Greenberg SM, Leder P. The IP-10 chemokine binds to a specific cell surface heparan sulfate site shared with platelet factor 4 and inhibits endothelial cell proliferation. J Exp Med 1995;182:219-231.
125. Angiolillo AL, Sgadari C, Taub DD, Liao F, Farber JM, Maheshwari S, Kleinman HK, Reaman GH, Tosato G. Human interferon- inducible protein 10 is a potent inhibitor of angiogenesis in vivo. J Exp Med 1995;182: 155-162.
126. Sun Y, Finger C, Alvarez-Vallina L, Cichutek K, Buchholz CJ. Chronic gene delivery of interferon-inducible protein 10 through replication-competent retrovirus vectors suppresses tumor growth. Cancer Gene Ther 2005; 12: 900-912.
127. Report of the National High Blood Pressure Education Program Working Group on High Blood Pressure in Pregnancy. Am J Obstet Gynecol 2000;183:S1-S22.
128. ACOG practice bulletin. Diagnosis and management of preeclampsia and eclampsia. Number 33, January 2002. Obstet Gynecol 2002;99:159-167.
129. Sibai BM, Ewell M, Levine RJ, Klebanoff MA, Esterlitz J, Catalane PM, Goldenberg RL, Joffe G. Risk factors associated with preeclampsia in healthy nulliparous women. The Calcium for Preeclampsia Prevention (CPEP) Study Group. Am J Obstet Gynecol 1997;177:1003-1010. 130. Alexander GR, Himes JH, Kaufman RB, Mor J, Kogan M. A United States national reference for fetal growth. Obstet Gynecol 1996;87:163-168.
131. Nagaoka K, Nojima H, Watanabe F, Chang KT, Christenson RK, Sakai S, Imakawa K. Regulation of blastocyst migration, apposition, and initial adhesion by a chemokine, interferon gamma-inducible protein 10 kDa (IP-10), during early gestation. J Biol Chem 2003;278: 29048-29056.
132. Hanna J, Goldman-Wohl D, Hamani Y, Avraham I, Greenfield C, Natanson-Yaron S, Prus D, Cohen-Daniel L, Arnon TI, Manaster I, et al. Decidual NK cells regulate key developmental processes at the human fetal-maternal interface. Nat Med 2006;12:1065-1074.
133. Sarris AH, Broxmeyer HE, Wirthmueller U, Karasawas N, Cooper S, Lu L, Krueger J, Ravetch JV. Human interferon-inducible protein 10: Expression and purification of recombinant protein demonstrate inhibition of early human hematopoietic progenitors. J Exp Med 1993;178:1127-1132.
134. Sasaki S, Yoneyama H, Suzuki K, Suriki H, Aiba T, Watanabe S, Kawauchi Y, Kawachi H, Shimizu F, Matsushima K, et al. Blockade of CXCL10 protects mice from acute colitis and enhances crypt cell survival. Eur J Immunol 2002;32:3197-3205.
135. Loetscher M, Gerber B, Loetscher P, Jones SA, Piali L, dark- Lewis I, Baggiolini M, Moser B. Chemokine receptor specific for IP10 and mig: Structure, function, and expression in activated T- lymphocytes. J Exp Med 1996; 184: 963-969.
136. Liu L, Callahan MK, Huang D, Ransohoff RM. Chemokine receptor CXCR3: An unexpected enigma. Curr Top Dev Biol 2005;68:149- 181.
137. Cole KE, Strick CA, Paradis TJ, Ogborne KT, Loetscher M, Gladue RP, Lin W, Boyd JG, Moser B, Wood DE, et al. Interferon- inducible T cell alpha chemoattractant (I-TAC): A novel non-ELR CXC chemokine with potent activity on activated T cells through selective high affinity binding to CXCR3. J Exp Med 1998;187:2009- 2121.
138. Cox MA, Jenh CH, Gonsiorek W, Fine J, Narula SK, Zavodny PJ, Hipkin RW. Human interferon-inducible 10-kDa protein and human interferon-inducible T cell alpha chemoattractant are allotopic ligands for human CXCR3: Differential binding to receptor states. Mol Pharmacol 2001; 59:707-715.
139. Sauty A, Colvin RA, Wagner L, Rochat S, Spertini F, Luster AD. CXCR3 internalization following T cell-endothelial cell contact: Preferential role of IFN-inducible T cell alpha chemoattractant (CXCL11). J Immunol 2001; 167:7084-7093.
140. Colvin RA, Campanella GS, Sun J, Luster AD. Intracellular domains of CXCR3 that mediate CXCL9, CXCL10, and CXCL11 function. J Biol Chem 2004;279:30219-30227.
141. Dajotoy T, Andersson P, Bjartell A, Lofdahl CG, Tapper H, Egesten A. Human eosinophils produce the T cell-attracting chemokines MIG and IP-10 upon stimulation with IFN-gamma. J Leukoc Biol 2004;76:685-691.
142. Hoffmann A, Levchenko A, Scott ML, Baltimore D. The IkappaB- NF-kappaB signaling module: Temporal control and selective gene activation. Science 2002;298:1241-1245.
143. Taub DD, Lloyd AR, Conlon K, Wang JM, Ortaldo JR, Harada A, Matsushima K, Kelvin DJ, Oppenheim JJ. Recombinant human interferon- inducible protein 10 is a chemoattractant for human monocytes and T lymphocytes and promotes T cell adhesion to endothelial cells. J Exp Med 1993;177:1809-1814.
144. Taub DD, Sayers TJ, Carter CR, Ortaldo JR. Alpha and beta chemokines induce NK cell migration and enhance NK-mediated cytolysis. J Immunol 1995;155:3877-3888.
145. Dewald B, Moser B, Barella L, Schumacher C, Baggiolini M, dark-Lewis I. IP-10, a gamma-interferoninducible protein related to interleukin-8, lacks neutrophil activating properties. Immunol Lett 1992;32:81-84.
146. Qin S, Rottman JB, Myers P, Kassam N, Weinblatt M, Loetscher M, Koch AE, Moser B, Mackay CR. The chemokine receptors CXCR3 and CCR5 mark subsets of T cells associated with certain inflammatory reactions. J Clin Invest 1998;101:746-754.
147. Muehlinghaus G, Cigliano L, Huehn S, Peddinghaus A, Leyendeckers H, Hauser AE, Hiepe F, Radbruch A, Arce S, Manz RA. Regulation of CXCR3 and CXCR4 expression during terminal differentiation of memory B cells into plasma cells. Blood 2005;105:3965-3971.
148. Farber JM. Mig and IP-10: CXC chemokines that target lymphocytes. J Leukoc Biol 1997;61:246-257.
149. Sallusto F, Lenig D, Mackay CR, Lanzavecchia A. Flexible programs of chemokine receptor expression on human polarized T helper 1 and 2 lymphocytes. J Exp Med 1998; 187:875-883.
150. Piali L, Weber C, LaRosa G, Mackay CR, Springer TA, dark- Lewis I, Moser B. The chemokine receptor CXCR3 mediates rapid and shear-resistant adhesion-induction of effector T lymphocytes by the chemokines IP10 and Mig. Eur J Immunol 1998;28:961-972.
151. Loetscher M, Loetscher P, Brass N, Meese E, Moser B. Lymphocyte-specific chemokine receptor CXCR3: Regulation, chemokine binding and gene localization. Eur J Immunol 1998;28:3696-3705.
152. Bonecchi R, Bianchi G, Bordignon PP, D’Ambrosio D, Lang R, Borsatti A, Sozzani S, Allavena P, Gray PA, Mantovani A, et al. Differential expression of chemokine receptors and chemotactic responsiveness of type 1 T helper cells (Th1s) and Th2s. J Exp Med 1998;187: 129-134.
153. Yamamoto J, Adachi Y, Onoue Y, Adachi YS, Okabe Y, Itazawa T, Toyoda M, Seki T, Morohashi M, Matsushima K, et al. Differential expression of the chemokine receptors by the Th1- and Th2-type effector populations within circulating CD4+ T cells. J Leukoc Biol 2000; 68:568-574.
154. Gangur V, Simons FE, Hayglass KT. Human IP-10 selectively promotes dominance of polyclonally activated and environmental antigen-driven IFN-gamma over IL-4 responses. FASEB J 199