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This article discusses a longitudinal study that examined the long-term effects of severe-to-critical SARS-CoV-2 infection on neutrophil activation. The study found that the release of neutrophil extracellular traps (NETs) was associated with the severity of pulmonary damage and other multi-visceral manifestations of the infection. The levels of NET components remained elevated 3 and 6 months after infection, suggesting ongoing neutrophil activation. The study also identified correlations between NET components and clinical or biological features of the infection. Further research is needed to determine the influence of NETosis on the development of respiratory sequelae. [Extracted from the article]

Summary: Background: Patients with extensive burns are at risk of developing candidemia. Objectives: To identify potentially modifiable risk factors and outcomes of candidemia in critically ill burns patients. Patients and Methods: Retrospective matched cohort study including adult burns patients. Patients who developed candidemia were matched with burns patients with Candida spp colonisation and sepsis or septic shock without candidemia in a ratio of 1:3 (same severity scores and colonisation index). Univariate and multiple regression analyses were performed. Results: Of 130 severely burned patients with Candida spp colonisation and at least one episode of sepsis or septic shock, 14 were diagnosed with candidemia. In the candidemia group, patients had a median (IQR) total burns surface area (TBSA) of 57 (38‐68)%, SAPSII of 43 (36‐58) and ABSI of 11 (8‐13). Multiple regression analysis showed that only duration of prior antibiotic therapy was independently associated with candidemia. ICU mortality was higher in the candidemia group (71% vs 35% [P = 0.02]). The log‐rank test for 28‐day mortality comparing patients with candidemia treated with an empirical strategy vs a curative strategy did not reach significance (P = 0.056). Conclusions: Burns patients having received recent antibiotherapy have a higher risk of candidemia. Antifungal strategies did not influence outcome in this series. [ABSTRACT FROM AUTHOR]

4‐Aminopyridine (4‐AP) is a recent treatment indicated to improve walking in patient with multiple sclerosis. We report the first case of pulmonary arterial hypertension (PAH) that we attribute to the use of 4‐AP. A 64‐year‐old woman with multiple sclerosis presented with dyspnea. After excluding other secondary causes of pulmonary hypertension, a diagnosis of severe PAH due to 4‐AP was made based on right heart catheterization. History revealed that the dyspnea began with the initiation of 4‐AP. After discontinuation of 4‐AP therapy and initiation of ambrisentan and tadalafil, dyspnea and pulmonary arterial pressure have improved significantly and one specific PAH treatment was stopped. 4‐AP is an outward rectifying potassium channel blocker with a vasoconstrictor effect in animal's pulmonary artery. According to the chronological sequence of events, the lack of other etiology, and its pharmacological plausibility, 4‐AP is highly suspected to have induced our patient's PAH. [ABSTRACT FROM AUTHOR]

Aim Leiomyomas, monoclonal tumors developed by the transformation of myometrium somatic stem cells, are a major health concern that can severely impair quality of life. Pathological alterations of signaling pathways have been recognized as a key feature in a variety of human diseases. Our objective was to analyze treatment with all-trans-retinoic acid (ATRA) by suppression of the phosphoinositide 3-kinase (PI3K) pathway on growth, signaling pattern and interactions among PI3K/B-cell lymphoma 2 (Bcl2)/retinol leiomyoma proteins. Methods Cultures of paired myometrium and leiomyoma cells from premenopausal women undergoing hysterectomy were collected. Western blot and analysis of variance were used for analysis. Results Significant differences were detected between treatment with ATRA alone or with LY294002 (a PI3K growth suppressor) in response to treatment and among cell samples and cell numbers. Leiomyoma cells were less affected. Immunochemical analysis of signaling patterns demonstrated that treatments affected most of the examined protein levels differently. Significant differences between the cell type responses to treatment in pyruvate phosphate dikinase 1 (pPDK1), Bad and pβ-catenin levels were identified. The pβ-catenin level showed highly significant interaction between response to treatment and cell type. Conclusions ATRA treatment on PI3K pathway suppression significantly affected growth, signaling pattern and interactions among PI3K/Bcl2/retinol proteins involved in the growth, survival and apoptosis of leiomyomas. Interpretation of our results suggests that increasing knowledge of the role of signaling interplay in the pathogenesis of leiomyomas may present an opportunity to use specific signal transduction inhibitors for treating and preventing this disorder. [ABSTRACT FROM AUTHOR]

Labeled problem Embryo implantation remains the main limiting factor in assisted reproductive medicine (20% success rate). Methods of study An endometrial immune profiling was performed among 394 women with the previous history of repeated embryo implantation failures (RIF). The endometrial immune profile documented the ratio of IL-15/Fn-14 mRNA as a biomarker of uNK cell activation/maturation (together with the uNK cell count) and the IL-18/TWEAK mRNA ratio as a biomarker of both angiogenesis and the Th1/Th2 balance. According to their profile, we recommended personalized care to counteract the documented dysregulation and assessed its effects by the live birth rate (LBR) for the next embryo transfer. Results Endometrial immune profiles appeared to be dysregulated in 81.7% of the RIF patients compared to control. Overactivation was diagnosed in 56.6% and low activation in 25%. The LBR among these dysregulated/treated patients at the first subsequent embryo transfer was 39.8%. Conclusion Endometrial immune profiling may improve our understanding of RIF and subsequent LBR if treated. [ABSTRACT FROM AUTHOR]

A catanionic mixture based on three components, N-dodecyldiethanolamine, decanoic acid and azelaic acid, was prepared and investigated from a physico-chemical point of view and for its antimicrobial activity. The spontaneously formed vesicles were characterized by dynamic light scattering, zeta potential and electron microscopy. These experiments showed spherical vesicles with an average diameter of 200 nm and a zeta potential of +22 mV. The antimicrobial activity of the vesicle dispersion against different strains ( Staphylococcus aureus, Escherichia coli, Staphylococcus epidermidis, Propionibacterium acnes and Candida albicans), was evaluated and compared with the activity of the components separately. The synergistic effect of these bioactive self-assembled catanionic vesicles on the spectrum of activity and on the cytoplasmic membrane is discussed. [ABSTRACT FROM AUTHOR]

Problem a2 isoform of vacuolar ATPase ( Atp6v0a2) is important for maintaining the delicate immunological balance required for successful pregnancy. The objective of this investigation is to study the dynamic changes in spleen and blood that appear during spontaneous abortion in mice. Method of study Atp6v0a2 was measured in multiple immune cell populations from spleen and blood recovered from non-abortion-prone and abortion-prone mating combinations. Results Atp6v0a2 expression was significantly lower ( P ≤ 0.01) in the spleen recovered from abortion-prone ♀ CBA × ♂ DBA mating on days 12 and 16 of pregnancy when compared to non-abortion-prone ♀ BALB/ c × ♂ BALB/c and ♀ CBA × ♂ BALB/c matings. Flow cytometric studies showed that significantly decreased expression of Atp6v0a2 in splenic CD4+, CD8+, CD19+, and CD14+ cells directly correlated with the high percentages of fetal resorption observed in abortion-prone mating on days 12 and 16 of pregnancy. In blood, CD4+, CD8+, and CD19+ cells had a significantly reduced expression of Atp6v0a2 in abortion-prone mating compared to the non-abortion-prone mating combinations only on day 12. Conclusion This deceased expression of Atp6v0a2 in the various immune cell populations of the spleen and blood suggests that the maternal environment is not supportive to fetus and leads to poor pregnancy outcome in the abortion-prone mating model. [ABSTRACT FROM AUTHOR]

The objective was to examine if IVF/ICSI repeated implantation failures (IF) or recurrent miscarriages (RM) could be related to preconceptional endometrial deregulations. IF was defined as the absence of pregnancy despite the transfer of at least ten IVF/ICSI good quality embryos, and RM as having at least three unexplained miscarriages. Fertile controls (FC) were women who had given birth at least once. Endometrial biopsy was performed in the mild luteal phase of a non-conceptual cycle (five women were selected in each group). Affymetrix chips (GeneChip Human Genome U133 Plus2.0 Array) were used for hybridization. Data were normalized by the gcRMA method, and raw p values adjusted by the Bonferroni procedure (1%). Differential expression of selected genes was analysed using real-time PCR. Gene networks and biological functions were explored using the Ingenuity Pathways Analysis software. Endometrial gene expression profiles at the time of uterine receptivity differ dramatically in the endometrium among FC, RM, and IF patients. Compared to FC, 2126 and 2477 genes are differentially expressed in IF and RM groups, respectively, and 2363 between IF and RM. In both conditions, differential gene expression referred mainly to DNA transcription and expression. Other main cellular functions deregulated in IF conditions correspond to cell morphology, cellular development, cell cycle, and cellular assembly, while in RM conditions, deregulated cellular functions relate to cell signalling (degradation of cyclic AMP and calcium metabolism) and cellular maintenance. In both conditions, there is an over-representation of deregulations related to the haematological system. In the IF condition, cell-mediated immune response and nervous system development and function are highly deregulated, while in RM patients, main deregulations are in organ and tissue development, humoral immune response, and muscular system development and function. Extensive endometrial deregulations are present before conception in patients who experienced IF or RM with both distinct and common deregulation. Copyright © 2011 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Background Patients with kidney failure frequently require the formation of an arterio-venous fistula (AVF) in which a vein is connected to an artery resulting in arterialization of the vein to allow adequate blood flow into an external 'artificial kidney'. In most patients, neo-intimal hyperplasia (NIH) ensues, causing narrowing and subsequent occlusion of the vein, leading to significant morbidity. The cellular events causing venous NIH may serve as ideal targets for molecular-based therapies. However, therapeutic gene delivery into the vascular system is seriously impeded by problems related to the low efficacy and toxicity of targeted viral vector delivery. Materials and methods To explore the feasibility of a plasmid-based vascular gene delivery system, we established a rat AVF model that develops NIH. Plasmids encoding for reporter or therapeutic genes were delivered into the blood vessels at the time or after AVF formation. Results Intra-luminal injection of plasmid into the AVF resulted in extensive and long-term reporter gene expression at the venous limb mainly at the site of NIH formation. Transgene expression was confined to endothelial cells and myofibroblasts that migrate inwards from the adventitia and form the NIH lesion. There was no detrimental tissue reaction to plasmid delivery, contrasting with the severe inflammatory response observed after adenovirus infection. Intra-vascular delivery of a plasmid carrying the endothelial nitric oxide synthase gene resulted in sustained production of nitric oxide, previously shown to mitigate NIH formation. Conclusions These findings open the possibility of vascular transduction with naked DNA bearing therapeutic genes in areas prone to NIH to ameliorate vein graft pathologies using simple and clinically applicable vector delivery methods. Copyright © 2011 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Abdominoperineal resection (APR) is the only curative treatment for recurrent or persisting squamous cell carcinoma of the anus after radiochemotherapy. A vertical rectus abdominis myocutaneous (VRAM) flap reduces perineal morbidity. The sexual life (SL) of women after APR is unknown. Aims of this study were to evaluate SF of women after APR. 47 women alive after APR performed between 1996 and 2007 were included. SL was evaluated using the female sexual function index (FSFI) score. 29 (62%) women answered the questionnaire: 15 (52%) had a VRAM and 16 (55%) a colpectomy. Among the 21 patients with SL before surgery, 16 (76%) still had intercourse with a mean FSFI score of 19.5 ± 10.9 [4.8-36]. Main difficulties reported were troubles of lubrification, orgasm, and dyspareunia. Confection of a VRAM did not influence the recovery of SL ( P = 0.717). Colpectomy reduced return of SL ( P = 0.026). Among women who had SL before APR, 76% still had sexual intercourse after. Colpectomy seems to reduce SL. [ABSTRACT FROM AUTHOR]

Citation Chaouat G, Petitbarat M, Dubanchet S, Rahmati M, Ledée N. Tolerance to the Foetal Allograft? Am J Reprod Immunol 2010 In this review, we will detail the concept of tolerance and its history in reproductive immunology. We will then consider whether it applies to the foetal–maternal relationship and discuss the mechanisms involved in non-rejection of the foeto-placental unit. [ABSTRACT FROM AUTHOR]

Citation Clark DA, Chaouat G, Wong K, Gorczynski RM, Kinsky R. Tolerance mechanisms in pregnancy: a reappraisal of the role of class I paternal MHC antigens. Am J Reprod Immunol 2010; 63: 93–103 Problem Allogeneic pregnancies have a survival advantage over syngeneic pregnancies, and paternal Class I MHC antigens have been implicated. In humans, HLA-C and HLA-G and E are expressed by subpopulations of fetal trophoblast. In mice, Qa-2, a Class Ib antigen, and classical H-2K antigens have been described. However, the mechanism of prevention of embryo demise in utero has not been critically assessed, and a number of conflicting ideas have not been addressed. The αβ T-cell receptor recognizes peptide bound to the groove in Class I MHC, and peptides have profound effects on the interaction of KIR receptors on T and NK cells with Class I MHC. Methods Data on prevention of pregnancy loss (abortion) in poly IC-treated mice were reviewed along with information about prevention of losses in the abortion-prone CBA × DBA/2 model. This information was combined with data on paternal antigen expression at different times in pregnancy when key events determining outcome are thought to transpire, and role of tolerance signaling molecules such as CD200. Current data on models supporting a role for ‘true’ uterine NK cells (TuNKs) versus blood NK cells in the uterus (BuNKs) and role of MHC–KIR interaction were reviewed along with incompatible data in the literature. Results Whilst paternal Class I MHC appears important, there is an important role for paternal non-MHC minor antigens (small peptides) that bind to the antigen-presenting groove of Class I MHC. BuNKs along with CD8+ T cells and Treg cells appear more important than TuNKs where the role of the latter appears primarily to promote angiogenesis. When during pregnancy the maternal immune system cells are first exposed to paternal Class I + peptide is uncertain, but at the time of implantation, if not earlier, seems likely. Conclusion Suppression of pregnancy loss by paternal/embryo Class I MHC depends on the presence of paternal peptides. This greatly complicates existing models of Class I–KIR interactions in feto-maternal tolerance or rejection. It is important to consider all the data when devising explanatory models. [ABSTRACT FROM AUTHOR]

Problem Previous data have shown that ‘danger’ signals, such as bacterial lipopolysaccharide (LPS) acting via toll-like receptors (TLR), are conditions antecedent to early pregnancy failure in several murine abortion models. Indeed, the abortion rate increased in the CBA × DBA/2 model after injection of tumor necrosis factor-α (TNF-α) + interferon (IFN-γ) on gestation day (GD) 7.5 only if the LPS–TLR signaling pathway was intact. High rates of cytokine-boosted abortion >80% loss can be demonstrable in certain animal colonies that have a high endogenous (spontaneous) rate of resorption (30–50%). A specific role for LPS on GD 0.5 determines the endogenous loss rate and on GD 6.5 the responsiveness to cytokine boosting of losses. Th1 cytokines (or the stress that induces these cytokines) increase intestinal permeability and absorption of luminal contents. It was predicted that intestinal availability of LPS was a major factor in the endogenous and cytokine-boosted resorption rates. Method of study A fixed weight of fresh mouse droppings from CBA/J female mice house in a high-abortion-rate (30–40%) colony at Clamart, France was homogenized and filtered. Fresh mouse droppings from a low-abortion-rate (10–15%) colony in Berlin, Germany were similarly processed. LPS was assayed using the Limulus amoebocyte lysate bioassay. Results To our surprise, there was no significant difference in LPS content of fecal samples from the two colonies. Conclusion A high endogenous rate of abortion and cytokine (or stress) boosted abortion in the CBA × DBA/2 model is not explained by the LPS content of feces. Possible explanations include: fecal LPS does not reflect small intestinal LPS, there are additional TLR signals besides LPS that are important and endogenous stress levels may be higher in high-abortion-rate colonies, so permeability of the intestine (and Th1 cytokine levels) may be already higher. These data have implications for studies on the role of flora in human pregnancy problems. [ABSTRACT FROM AUTHOR]

Problem Spontaneous abortions in the CBA × DBA/2 model are normally reported as number of resorptions/total number of implantations (R/T), pooling data from individual mice. The significance of differences between groups has been determined using non-parametric statistics (e.g. chi-square or Fisher’s Exact test) based on a priori predictions. Recently, it has been argued that medians with box plots should replace the accepted standard, but this deprives readers of data needed to verify P-values, and leads to inferences incompatible with biological and statistical reality. Method of study Raw data on 173 individual CBA × DBA/2 matings were analyzed by median and mean, along with R/T data from 18 independent experiments containing 5–10 mice per group. Raw data from 19 CBA × BALB/c matings were similarly analyzed. Results Individual CBA × DBA/2 mouse resorption rates showed a non-Gaussian distribution, but the mean and median differed by <0.5%. Resorption data from 6 and 12 independent pools of mice were normally distributed. Only the mean enabled a between-group P-value calculation. CBA × BALB/c matings gave a median of 0 and mean of 5.1%; the data were not normally distributed, but that was because of a bimodal distribution. One group of mice had 0 abortions, and the second a mean of 13.9% abortions, and the data from the latter group were normally distributed. Conclusion Although it is possible to compare individual mice, and even individual implantation sites, in resorption (abortion) studies, as the relevant question is the significance of differences between treatment groups of mice, and reproducibility, the established classical method of reporting R/T should continue to be provided. In CBA × BALB/c matings, where abortion rates are low, using the median is misleading and may obscure the existence of two distinct populations. [ABSTRACT FROM AUTHOR]

Problem An important subset of implantation defects/early abortion seems to be linked with a deregulation of the interleukin (IL)-12/IL-15/IL-18 system as well as tumor necrosis factor (TNF)- and natural killer (NK)-controlled/mediated networks at the decidual placental interface, both in case of deficient or excess expression. The presence of TNF in high amounts in the pre/peri-implantation uterus and its pivotal role during pregnancy are difficult to reconcile with its abortive effects in ongoing pregnancy. We therefore searched for regulators of the IL-12/IL-18 family of cytokines as well as for antagonist(s) of TNF with potentially selective effects on implantation. Method of study We first used Swiss mice to verify the presence in the murine reproductive tract of ‘new members’ of the IL-12 family of cytokines, IL-23 and IL-27, as well as of tumor necrosis factor-like WEAK inducer of apoptosis (TWEAK), described as acting with TNF as Yin and Yang of innate immunity in murine placenta/ decidua at days 0–12.5. We then compared expression by RT-PCR in the CBA × DBA/2, and CBA × BALB/c murine mating combinations. Finally, we performed in vivo neutralization experiments of TWEAK and IL-27. Results Immunohistochemistry (IHC) studies showed that IL-23, IL-27, and TWEAK were expressed at the interface. For RT-PCR, IL-23 expression peaked at day 9.5 in the non-aborting mating combination, a peak absent in the aborting one, and thus difficult to explain except by invoking a feed back on EB13 (Epstein–Barr virus-induced gene 3 cytokine). Most important, an immediate post-mating IL-27 hyper expression was seen in the CBA × DBA/2 mating compared to CBA × BALB/c one. The difference in expression resurged and was statistically very significant by days 6.5–9.5, compatible with an early activation of inflammation on day 0.5 which would then peak again in the ‘resorption window’ where takes place the early NK/mph activation described by Baines et al. A significant TWEAK expression was present in both strains from days 0.5 to 4.5 peaking in both cases in the first days when it is known that intra uterine TNF also reaches high levels as a component of post-mating inflammation. However, it was lower from day 1.5 in the abortion-prone CBA × DBA/2 mating combination, and almost absent by days 6.5–9.5 when compared to the non-aborting CBA × BALB/c mating combination. In both mating combinations, neutralization of TWEAK-enhanced resorption rates, but surprisingly so did IL-27 neutralization. Conclusion TWEAK is likely offering protection against the deleterious effects of TNF in implantation explaining embryo survival in a TNF-rich environment, and equal number of implants in both strains. However, there is a clear difference of protection in abortion-prone mating peaking in the abortion/resorption window but starting early, and therefore possible links with the prevention of abortion in CBA × DBA/2 matings by interfering with complement activation as recently described by Girardi et al. are discussed, as well as consequences for our current view of feto-maternal ‘seed and soil’ interplay. The apparently paradoxical effects of IL-27 neutralization are also discussed. [ABSTRACT FROM AUTHOR]

Problem To evaluate the influence of ovarian steroids on IL-18, IL-15 and angiopoietin-2 mRNA expression in the endometrium in the mid luteal phase. Method of study We quantified IL-18/GAPDH, IL-18 BP/GAPDH, IL-15/GAPDH and angiopoietin-2/GAPDH in the endometrium by quantitative polymerase chain reaction on day 21 of the cycle. We first compared cytokines expression over two natural cycles ( n = 15) then between natural and oestrogen–progestin replacement treatment ( n = 18). Results Endometrial IL-18, IL-18 BP, IL-15 and angiopoietin-2 mRNA expression did not change over two natural cycles. Addition of exogenous hormones significantly decreased IL-18 and IL-18 BP mRNA expression but not influence IL-15 or angiopoietin-2 ratios. This was also observed with immunohistochemistry. Conclusion Exogenous oestro-progestative hormones influence endometrial IL-18 system expression involved in angiogenesis and in the uterine natural killer (uNK) cell activation pathway during the implantation process. [ABSTRACT FROM AUTHOR]

Chaouat G, Ledée-Bataille N, Dubanchet S. Immunological similarities between implantation and pre-eclampsia. AJRI 2005; 53:222–229© Blackwell Munksgaard, 2005Cytokines are involved in implantation success and failure. We envisage that they could be similarly involved in pre-eclampsia (PE).First, we review the primipaternity and primiparity concepts and then why natural killer (NK) cells are involved in implantation. We stress that the common event in all PE is vascular remodelling.We conclude that PE could involve cytokine and/or NK dysfunctions, and propose a working hypothesis. [ABSTRACT FROM AUTHOR]

PROBLEM: Prevention of spontaneous abortion by allogeneic mononuclear leukocyte immunotherapy has proven ineffective in the CBA x DBA/2 murine abortion model if the leukocytes are stored overnight before inoculation. The mechanism and generality of the phenomenon has not been elucidated. METHODS: As prevention of recurrent abortion in the CBA x DBA/2 model requires allogeneic BALB/c lymphoid cells bearing paternal antigens and the tolerance-signaling molecule CD200 (OX-2), we evaluated effects of cell storage on cell surface CD200 expression using flow cytometry of fresh or stored cells stained with monoclonal anti-CD200 antibody. Release of putative CD200 molecules into culture supernatant during storage was tested by the ability of supernatants to block binding of anti-CD200 to freshly isolated cells. Similar studies were done using human peripheral blood mononuclear leukocytes. Possible binding of soluble CD200 to immunoglobulin G (IgG) molecules in plasma as a basis for the anti-abortive effect of intravenous immunoglobulin G (IVIG) was tested using the standard peripheral blood lymphocyte (PBL) natural killer (NK) cell lysis of 51Cr-labeled K562 cells and monoclonal anti-human CD200 antibodies. RESULTS: Loss of anti-abortive effect of BALB/c cells with overnight storage at 4°C and blocking of protective effect of freshly isolated cells with anti-CD200 antibody was confirmed. Supernatants of stored cells acquired a low level of protective activity against abortion in the CBA x DBA/2 model. Cell surface CD200 was lost with overnight storage at 4 or 22 C, and supernatants acquired the ability to block binding of anti-CD200 antibody to fresh cells. Similar results were obtained using human PBL. However, if cells were stored overnight in IgG containing plasma, binding was not blocked. Suppression of NK cell lysis by PBL was abrogated if anti-CD200 antibody was added to the assay. CONCLUSIONS: Loss of the tolerance signal CD200 from allogeneic cel... [ABSTRACT FROM AUTHOR]

PROBLEM: Previous data have shown 'danger' signals, such as bacterial lipopolysaccharide (LPS) acting via toll-like (tlr) receptors are required for early pregnancy failure in several murine abortion models. Indeed, the abortion rate increased in the CBA x DBA/2 model after a gestation day (gd) 7.5 injection of tumour necrosis factor (TNF)-α +interferon (IFN)- γ only if the LPS-tlr signalling pathway was intact. High rates of cytokine- boosted abortion > 80%loss can be achieved in certain animal colonies, that have a high endogenous (spontaneous) rate of resorption (30-50%). A specific role for LPS has been postulated to determine both the endogenous and cytokine-boosted losses. METHODS: To test the role of LPS in spontaneous and cytokine-boosted abortions, recombinant TNF-α +IFN-γ and LPS were injected in different doses and sequences intraperitoneally (i.p.) into CBA-DBA/2 mated mice in the Toronto General Research Institute animal facility where the endogenous abortion rate is < 30%.The effects of poly IC,a tlr3 agonist that induces IFN-γ that can reverse LPS-induced tolerance,and effects of anti-MD-1 on TNF-α induction by LPS, poly IC, CPG, or HSP in vitro were also examined. RESULTS: A high endogenous rate of loss similar to that seen in Clamart could be achieved by increasing exposure to LPS on the morning after mating (gd 0.5). The magnitude by which the abortion rate could be increased by an i.p. injection of 2000 u TNF-α +1000 u IFN-γ on gd 7.5 was independent of the endogenous rate of loss, and could not be increased by doubling the dose. One microgram of LPS given on day 7.5 achieved a similar rate of loss, and if given with the cytokines, synergistically boosted the rate of loss to near Clamart rates. LPS given 1 day prior to the cytokines abrogated the cytokine effect, whereas LPS given day 0.5 had no significant effect on the response to day 7.5 cytokine injection. Blocking MD-1 inhibited TNF-&... [ABSTRACT FROM AUTHOR]

PROBLEM: In the human, lymphocyte alloimmunization immunotherapy (IT)has been proposed as a treatment for recurrent spontaneous abortion (RSA). This treatment has been the subject of debate for a long period of time. Recently, it has been proposed to extend such a treatment for implantation failure in humans, and I was asked to express my opinion on this topic. METHODS: I reviewed the evolution and theories and current paradigms in reproductive Immunology and rationales proposed for IT. New discoveries show the complexity of implantation as a step by step developmental event, in mice and humans, and as such led me to re-examine paradigms currently evoked for extension of IT to implantation. Such a re-examination obviously leads me to re-question the basis of IT for SA itself. CONCLUSIONS: I conclude that the Th1/Th2 paradigm, evoked as the current basis for IT, and as useful as it has been to explain pregnancy, is no longer sufficient. It is especially insufficient to explain the process of implantation, which involves inflammatory molecules and cannot fit in such a scheme. It ensues that alloimmunization has no scientific rationale for the treatment of human implantation as a whole, and should not be applied broadly at such a stage of pregnancy. Furthermore, its use in RSA should be re-examined. [ABSTRACT FROM AUTHOR]

PROBLEM: The aim of the present study was to define the expression of CXCR4 and CCR5 on non-cultured non-stimulated primary human trophoblast cells (TCs) immediately after their immunopurification. METHOD OF STUDY: We have evaluated by flow cytometric analysis and immunofluorescence, highly purified primary TCs prepared from first trimester (8.2 ± 0.3 weeks, n = 15) and term (Caesarean section, n = 10) placentae for the cell surface and intracellular expression of CXCR4 and CCR5. RESULTS: There was a high level of individual variability for CXCR4 and CCR5 expression between trophoblast batches. In first trimester and term placentae TCs, we found a greater number of TCs preparations expressing intracellular CXCR4 than CCR5 (P < 0.05). Both receptors were predominantly localized in the intracellular compartment of TCs, whatever if isolated from first trimester or term placentae. CONCLUSIONS: The functional consequences of the predominance of CXCR4 expression and of cellular addressing are briefly discussed. [ABSTRACT FROM AUTHOR]

Problem: Human in vitro fertilization (IVF) embryo transfer is accompanied by a low implantation rate even after a very successful IVF, and there are a certain number of ‘idiopathic sterilities’ which are due to repeated implantation failures. In the very same vein, the question of improving implantation rates is of prime importance in agricultural research to improve the management of livestock. Pre-implantation prenatal diagnosis cannot be accomplished in individuals who have a high rate of implantation failure, whether women undergoing IVF, or animals, during genetic cloning. Implantation cytokine networks need to be known in such a perspective. Methods: We review the evolution and theories in reproductive immunology, briefly deal with the complexity of implantation as a step by step developmental event, and then present some of our recent data in mice and human. Conclusions: We conclude that the T helper cell type 1/2 (Th1/Th2) paradigm, as useful as it has been to explain pregnancy, is no longer sufficient in view of the emerging complexity of the cytokine network at the materno-fetal interface. This is peculiarly true for implantation, which, as a step by step developmentally regulated process, involving inflammatory molecules, cannot fit into such a scheme. [ABSTRACT FROM AUTHOR]

Estrogens promote cell proliferation in normal and transformed mammary epithelial cells by inducing expression of hormone-responsive genes involved in the cell cycle. The action of antiestrogens is therefore central in regard to their potent inhibitory effects on estrogen-induced cell growth. We used normal human epithelial breast cells from primary cultures (HBE cells) to study hormonal (estrogen and antiestrogen) regulation on 3 key proteins involved in the apoptotic process: Bcl-2, p53 and caspase-3. The mammary adenocarcinoma cell line, MCF-7, was also used to study the molecular regulation of Bcl-2. In both HBE and MCF-7 cells, we found that estradiol (E2) induced an increase in Bcl-2 mRNA levels. This effect was counteracted in the presence of a pure antiestrogen, ICI 182780 (ICI). Alone, ICI did not modify either the Bcl-2 protein or mRNA levels in HBE cells, whereas in MCF-7, a strong downregulation of Bcl-2 mRNA was observed. In parallel, in HBE cells, we observed that E2 caused a decrease in p53 and caspase-3 protein levels, whereas ICI alone increased p53 and caspase-3 protein levels. The ICI effects on p53 and caspase-3 were partially counteracted by E2. Under the same experimental conditions, ICI exerts a potent pro-apoptotic effect, which was not counteracted by E2. In contrast, 4-hydroxytamoxifen was slightly weaker as a pro-apoptotic agent in HBE cells and its effects were reversed by E2. We demonstrate that in HBE cells, ICI reverses the anti-apoptotic action of E2 and alone acts as a highly potent pro-apoptotic molecule. These results provide new insight into treatment for breast cancer prevention. © 2003 Wiley-Liss, Inc. [ABSTRACT FROM AUTHOR]

PROBLEM: To determine if interleukin-16 (IL-16), IL-17, and IL-18 are present at the murine fetomaternal interface during pregnancy as a first step towards investigating their roles in fetomaternal relationship. METHODS: Expression of IL-16, IL-17, and IL-18, was assessed by immunohistochemistry (IHC) in the BALB/c x BALB/k (H2[supd] x H2[supk]), and the CBA/J x BALB/c non-abortion prone, and CBA/J x DBA/2 abortion prone matings. Enzyme-linked immunosorbent assay (ELISA) were performed for the two latter cytokines to compare local production in the abortion prone CBA/J x DBA/2 versus the nonabortion prone CBA/J x BALB/c matings. RESULTS: Expression of IL-17 was borderline. The anti-IL-16 staining specifically localized in the uterine stroma and glandular epithelium and was rather low in the placenta. IL-18 staining started in the peri-implantation uterus in the basal proliferative stroma, and was also traced, although weaker, in the glandular epithelium. In the immediate post-implantation period, a weak stromal staining persisted but there was a strong labeling of the ectoplacental cone. Interestingly, when the ectoplacental cone differentiates into placenta having a major histocompatibility complex (MHC) class I + spongiotrophoblast and a (MHC class I) labyrinth, a very strong transient labeling of uterine natural killer (u-NK) cells was found. Later in gestation, IL-18 was also produced by giant cell and spongiotrophoblast. Finally, we compared by ELISA the production of IL-17/-18 in CBA/J x DBA/2 and CBA/J x BALB/c matings. We detected significantly more IL-18 in the non-abortion prone combination decidua or placenta. CONCLUSION: The... [ABSTRACT FROM AUTHOR]

Different HLA-G monoclonal antibodies (mAbs) were first evaluated for their capability to identify soluble HLA-G (sHLA-G) in ELISA. Three of them, namely 87G, BFL.1 and MEM-G/9, when used as coating mAbs together with W6/32 capture mAb, identified β2-microglobulin (β2m)-associated-sHLA-G but not soluble HLA-B7 (sHLA-B7) in cell culture supernatants from transfected cells. By comparison, the anti-HLA class I mAb 90 did recognize both sHLA-G and sHLA-B7. By using these HLA-G mAbs, sHLA-G was identified in amniotic fluids as well as in culture supernatants of first trimester and term placental explants but not in cord blood. Intron 4-retaining sHLA-G isoforms were identified in some amniotic fluids by the use of an intron 4-specific mAb (16G1). Reactivity of these different HLA-G mAbs was then compared to determine their respective binding sites on soluble and membrane-bound HLA-G. Using both ELISA and flow cytometry analysis, we showed that they did not compete with each other, which suggested that they did not recognize the same determinants. Finally, we report that two mAbs directed against the α1 domain of HLA class I heavy chain (mAb 90 and YTH 862) did compete with 87G, therefore demonstrating that this latter mAb recognized an epitope localized on this external domain of HLA-G. [ABSTRACT FROM AUTHOR]

Culture supernatants of BW 5147 cells widely used for T-cell hybridization often manifest non-MHC-restricted, non-antigen-specific regulatory activities on the mixed lymphocyte reaction (MLR) of mouse cells. This report demonstrates that, whereas supernatants of BW 5147 cells grown at low concentrations (2 × 105/ml) enhanced MLR, high cell concentration (2 × 106/ml) supernatants markedly inhibited this reaction. BW 5147 cell-free extracts significantly inhibited MLR and in vitro antibody production (PFC), as well as the mitogenic response to lipopolysaccharide E. coil (LPS) of mouse spleen cells, but did not affect the response to an optimal dose of phytohaemagglutinin (PHA). Both supernatant and cell-free extract inhibitory activities were located in 60,000 MW fraction. Inhibitory material of low MW (< 12,000) was also found in high cell concentration supernatants. A similar suppressive activity was exerted by cell-free extracts of P3 × 63 NS cells used for B-cell hybridization. The suppressive activity seemed to stem from some kind of interaction between BW 5147 cells and the fetal calf serum (FCS) of the culture medium. Supernatants from subclones of BW 5147 cells obtained in selected batches of FCS and mains tamed in the same serum, even at high cell concentrations, did not affect MLR, whereas the supernatants from the same subclones maintained in other batches definitely suppressed this reaction. Thus, provided that culture conditions are chosen carefully, subclones of BW 5147 devoid of effect on in vitro immune reactions can be obtained. [ABSTRACT FROM AUTHOR]

The properties of mitomycin-treated and untreated spleen cells from allopregnant mice (bearing allogeneic foetuses) have been compared for suppressive activity of mixed lymphocyte reaction (MLR) of maternal strain spleen cells responding to paternal (or genetically related) strain cells. Mitomycin-treated cells in 1% normal mouse serum suppress an MLR if added at its onset while they cannot alter an already set up (day 2) MLR. Conversely untreated spleen cells are unable to affect an MLR if added at its onset while they suppress it when added on day 2 of an ongoing MLR. The two systems also differ in their m vitro retriggering requirements at the target cell (stimulator) level. Whereas the untreated, late acting population, in a k/d or k/a system, requires the contact with molecules coded by ‘S-G’ (or IC + S G) subregion, also present in the father's strain, the mitomycin-treated, early acting population, tested on similar panel does first appear non-specific. Suggestive evidence, however, is given that this apparent non-specificity may in fact veil clones of cells with specific recognition of ‘private’ Ia determinants. It is suggested that allopregnant mice elaborate two T suppressor cells, acting at different steps of the MLR. The usefulness of the first set (mitomycin-resistant suppression) for exploring the Ia chart is pointed out. [ABSTRACT FROM AUTHOR]

Adding mitomycin-treated spleen cells from an anopregnant mouse to a maternal-strain cells v, paternal-strain cells MLR was previously shown to decrease the intensity of the reaction, due to suppressor T cells. In the present study, spleen cells from allogeneically pregnant mice, were added without treatment at day 2 of an MLR of maternal strain responder cells raised against paternal-strain stimulators. It was shown that, while, on 2 day duration culture versus the same stimulator cells these cells are hyper-reactive compared to controls, yet they lead to a M LR of lower intensity than when the same operation was performed with control cells. This suppressive effect is T-cell dependent and use of high dilutions of anti-Thy 1+C suggests that the suppressor cell has a high density of Thy 1 surface antigen. Involvement of these cells in negative regulation of the late phase of the MLR is suggested. MLRs with cells from allopregnant mice as responders were themselves susceptible to this suppression and even more so than cells from virgin mice, indicating a possible physiological role. [ABSTRACT FROM AUTHOR]

The intensity of reticuloendothelial blockade by carrageenan, silica and ethyl stearate was measured and its effect studied on the susceptibility to tolerance induction by polysaccharide antigens in mice. The most intense RE depression reduced the tolerance threshold dose of levan only by 3-fold and that of dextran B512 not at all. The genetic resistance of BALB/c mice to tolerization with the α-3 glucose epitope of dextran B1355 was not overcome by carrageenan blockade which did, however, render them normally susceptible to tolerance induction by human γ-globulin. PFC responses to immunization by the polysaccharides were diminished by blockade, relative to its intensity and to the antigen itself. These and other data suggest that severe RE blockade (a) can promote tolerance by suppressing the active role(s) of the macrophages in immune induction rather than by sustaining circulating antigen, and (b) depresses responses to thymus-independent antigens, probably by an immunosuppressive influence mediated by damaged macrophages. [ABSTRACT FROM AUTHOR]

Tyrosine kinase inhibitors of the tyrphostin family which block EGF receptor kinase are reported to arrest the growth of psoriatic kera-tinocytes in vitro. Three tyrphostins with the potency ratio AG555 >> AG18 >> AG814 were found to arrest growth with no adverse cytotoxic effects. The potency ratio to inhibit keratinocyte proliferation follows their potency to inhibit EGF receptor kinase activity in vitro. These compounds represent novel leads for the therapy of psoriasis. [ABSTRACT FROM AUTHOR]

The article presents an obituary for immunologist Radslav Kinsky.

The author comments on the manuscript "Treatment with tumor necrosis factor (TNF) inhibitors and intravenous immunoglobulin (IVIG) improves live birth rates in women with recurrent spontaneous abortion (RSA)," by Edward E. Winger and Jane L. Reed. According to the author, the manuscript provides the views of the Alan Beer Reproductive centres and is welcomed because it deals with IVIG, not lymphocyte allo-immunization as a technique for treatment of recurrent pregnancy loss.