Fawaz Azizieh

Elevated levels of IL-8 in dengue hemorrhagic fever.

Dengue virus causes dengue fever, a mild febrile illness, and at times dengue hemorrhagic fever (DHF), a severe illness the pathogenesis of which is not fully understood. Given the crucial roles played by interleukin‐8 (IL‐8) as a chemoattractant cytokine and in inflammatory processes, levels of circulating IL‐8 in the sera and IL‐8 mRNA in the peripheral blood mononuclear cells (PBMC) were measured in 99 patients of a recent dengue epidemic that occurred in India in 1996 and in 21 normal healthy controls. Twenty‐six of the patients had dengue fever (DF) and the remaining 73 were diagnosed as having different grades of DHF. All the control normal sera were negative for IL‐8, so were their PBMC for IL‐8 mRNA. Increased levels of IL‐8 in the sera and IL‐8 mRNA in their PBMC were observed in patients with severe illness of DHF grades III and IV. Only two out of 26 patients of DF and one out of 10 DHF grade I patient were positive for IL‐8 and all three deteriorated to DHF grade IV within 24 hr. All six patients of DHF grade IV who died had higher serum level of IL‐8 above 200 pg/ml, the highest being 5,568 pg/ml in one patient; the presence of mRNA for IL‐8 was very high in all patients. A striking correlation was observed between increased levels of IL‐8 and severe DHF, with greater levels in patients with increased grade of the disease and death. These results suggest that IL‐8 may have an important role and may be an indicator of increasing severity of the disease and death. J. Med. Virol. 56:280–285, 1998.

Increased expression of pro-inflammatory cytokines in placentas of women undergoing spontaneous preterm delivery or premature rupture of membranes

Problem:  The objective of this study was to determine the levels of cytokines in the placentas of women undergoing preterm delivery (PTD) or premature rupture of membranes (PROM) as compared with women undergoing normal delivery at term.

Sequential production of cytokines by dengue virus-infected human peripheral blood leukocyte cultures

The study was undertaken to elucidate the sequence of appearance of T helper (Th)1‐ and Th2‐type cytokines in human peripheral blood leucocyte cultures infected in vitro with dengue type 2 virus. Commercial sandwich enzyme‐linked immunosorbent assay kits were used to assay the levels of tumour necrosis factor‐alpha (TNF‐α), interferon‐gamma (IFN‐γ), interleukin (IL)‐2, IL‐4, IL‐5, IL‐6, and IL‐10 in culture supernatants. Culture supernatants were also screened for the cytotoxic factor and the dengue virus titres determined. The cytokines that appeared in the culture supernatants on the first day post‐infection (p.i.) were cytotoxic factor, TNF‐α, IL‐2, and IL‐6; their levels were highest on the second day p.i. IFN‐γ appeared on the second day with a peak on the third day p.i. The levels of these cytokines declined quickly, except for human cytotoxic factor (hCF) and IL‐2. The cytokines that appeared later were IL‐10 and IL‐5 on the fourth day and IL‐4 on the sixth day p.i. Dengue virus replicated in the peripheral blood leucocyte (PBL) cultures and was present throughout the course of the study. The findings of the present study show that dengue virus induced a predominant Th1‐type cytokine response during the first 3 days of infection of PBL cultures that was replaced by a Th2‐type response later. J. Med. Virol. 59:335–340, 1999.

Modulation of cytokine production by dydrogesterone in lymphocytes from women with recurrent miscarriage

Objective  To examine the effects of dydrogesterone on the production of Th1 and Th2 cytokines by lymphocytes from women undergoing unexplained recurrent spontaneous miscarriage (RSM).

Pro‐inflammatory Maternal Cytokine Profile in Preterm Delivery

PROBLEM:  The objective of this study was to determine the levels of cytokines produced by maternal peripheral blood mononuclear cells (PBMC) upon stimulation with a mitogen, with autologous placental cells and with a trophoblast antigen extract.

Mitogen‐Induced Cytokine Responses of Maternal Peripheral Blood Lymphocytes Indicate a Differential Th‐Type Bias in Normal Pregnancy and Pregnancy Failure

Makhseed M, Raghupathy R, Azizieh F, Al‐Azemi MMK, Hassan NA, Bandar A. Mitogen‐induced cytokine responses of maternal peripheral blood lymphocytes indicate a differential Th‐type bias in normal pregnancy and pregnancy failure. AJRI 1999; 42:273–281

Maternal Cytokine Production Patterns in Women with Pre-eclampsia

Problem:  To determine the levels of cytokines produced upon mitogenic or antigenic stimulation of maternal peripheral blood mononuclear cells (PBMC) from women with pre‐eclampsia.

Th1/Th2 cytokine patterns and clinical profiles during and after pregnancy in women with multiple sclerosis

Pregnancy in multiple sclerosis (MS) patients is associated with a lower risk of progression and lower rate of exacerbation. These beneficial effects are reversed postpartum. Considering that the pathogenesis of MS appears to involve cell-mediated immune reactivity, and that pregnancy is accompanied by a depressed cell-mediated immunity, it has been proposed that the lower relapse rate and risk of progression of MS during pregnancy may be due to a pregnancy-associated down-regulation of cell-mediated immunity. In addition, pregnancy results in a shift towards a T helper (Th) 2 cytokine profile, which is presumably protective for MS. This study was aimed at investigating the relationship between clinical status of MS and cytokine levels in eight patients with MS who were followed through pregnancy and after delivery. Peripheral blood lymphocytes from these women were stimulated with a mitogen at different time points during and after gestation and the levels of Th1 cytokines (IFNgamma, TNFalpha) and Th2 cytokines (IL-4, IL-10) were estimated by ELISA. It was established that six of the eight MS patients studied showed a distinct shift from a Th2 cytokine bias during pregnancy towards a Th1 cytokine bias after delivery. These results suggest a possible association between decreased incidence of exacerbation of MS in pregnancy and a pregnancy-induced shift towards Th2 cytokine bias.

Progesterone-induced blocking factor (PIBF) modulates cytokine production by lymphocytes from women with recurrent miscarriage or preterm delivery.

Spontaneous miscarriage and preterm delivery are common complications of pregnancy. Pro-inflammatory cytokines have been shown to be associated with recurrent spontaneous miscarriage (RSM) and preterm delivery (PTD) and these have led to exploration of ways to downregulate pro-inflammatory cytokines and/or to upregulate anti-inflammatory cytokines. Progesterone-induced blocking factor (PIBF) is a molecule with inhibitory effects on cell-mediated immune reactions. We have ascertained the effects of PIBF on secretion of selected type 1 and type 2 cytokines by peripheral blood mononuclear cells from healthy non-pregnant women, women undergoing normal pregnancy, women with unexplained RSM and women with PTD. Peripheral blood mononuclear cells from 30 women with a history of unexplained RSM, 18 women undergoing PTD, 11 women with normal pregnancy and 13 non-pregnant healthy women were stimulated with a mitogen in the absence and presence of PIBF after which the levels of cytokines released into culture supernatants were determined by ELISA. Production of the type 2 cytokines IL-4, IL-6 and IL-10 by lymphocytes from the RSM and PTD groups and of IL-4 and IL-10 by lymphocytes from healthy pregnant women was significantly increased upon exposure to PIBF, while the levels of type 1 cytokines were not affected. Ratios of type 1:type 2 cytokines were decreased, suggesting a shift towards Th2 bias. PIBF did not affect cytokine production by lymphocytes from non-pregnant women. Thus, PIBF acts on lymphocytes in pregnancy to induce a type 1 to type 2 cytokine shift by upregulating the production of type 2 cytokines.

Intrauterine Growth Restriction: Cytokine Profiles of Trophoblast Antigen-Stimulated Maternal Lymphocytes

Intrauterine growth restriction (IUGR) is an important perinatal syndrome that poses several serious short- and long-term effects. We studied cytokine production by maternal peripheral blood lymphocytes stimulated by trophoblast antigens. 36 women with a diagnosis of IUGR and 22 healthy women with normal fetal growth were inducted. Peripheral blood mononuclear cells were stimulated with trophoblast antigens and levels of the proinflammatory cytokines IL-6, IL-8, IL-12, IL-23, IFNγ, and TNFα and the anti-inflammatory cytokines IL-4, IL-10, and IL-13 were measured in culture supernatants by ELISA. IL-8 was produced at higher levels by blood cells of the IUGR group than normal pregnant women, while IL-13 was produced at lower levels. IL-8, IFNγ, and TNFα were higher in IUGR with placental insufficiency than in normal pregnancy. IL-12 levels were higher and IL-10 levels were lower in IUGR with placental insufficiency than in IUGR without placental insufficiency. We suggest that a stronger pro-inflammatory bias exists in IUGR as compared to normal pregnancy and in IUGR with placental insufficiency when compared to IUGR without placental insufficiency. Several ratios of proinflammatory to anti-inflammatory cytokines also support the existence of an inflammatory bias in IUGR.