Novel HLA-G-binding leukocyte immunoglobulin-like receptor (LILR) expression patterns in human placentas and umbilical cords.

Human placentas are sources of cytokines, hormones and other substances that program receptive cells. One of these substances is HLA-G, which influences the functioning of both leukocytes and endothelial cells. In this study, we investigated the possibility that these and/or other types of cells in extraembryonic fetal tissues might respond to HLA-G by interacting with one or another of the leukocyte immunoglobulin-like receptors (LILR). LILRB1 is expressed by most leukocytes and LILRB2 is expressed primarily by monocytes, macrophages and dendritic cells. Analysis of term placentas by immunohistochemistry and Real Time PCR demonstrated that LILRB1 and LILRB2 protein and specific messages are produced in the mesenchyme of term villous placenta but are differently localized. LILRB1 was abundant in stromal cells and LILRB2 was prominent perivascularly. Neither receptor was identified in trophoblast. Further investigation using double label immunofluorescence indicated that placental vascular smooth muscle but not endothelia exhibit LILRB2. Term umbilical cord exhibited the same LILRB2 patterns as term placenta. Samples obtained by laser capture dissection of vascular smooth muscle in umbilical cords demonstrated LILRB2 mRNA, and double label immunofluorescence showed that cord vascular smooth muscle but not endothelium exhibited LILRB2 protein. The presence of LILRB1 in placental stromal cells and LILRB2 in vascular smooth muscle strongly suggest that HLA-G has novel functions in these tissues that could include regulation of placental immunity as well as development and function of the extraembryonic vasculature.

A commentary on gestational programming and functions of HLA-G in pregnancy.

HLA-G is an HLA class Ib gene that is highly expressed in human trophoblast cells. The single HLA-G mRNA is alternatively spliced to generate at least seven transcripts, three of which encode soluble isoforms. Many studies have shown that high levels of soluble antigens are associated with successful implantation and graft acceptance. To study expression, regulation and functions of two of the soluble isoforms, HLA-G5 and HLA-G6, we generated recombinant proteins in eukaryotic cells and developed monoclonal antibodies specific for each of the two proteins. In addition, we investigated the olive baboon Paan-AG gene as a potential functional correlate of HLA-G. Here, we present summaries of the studies that have been conducted in our laboratory using these tools and discuss the results within the context of the research on this topic that is ongoing in ours and other laboratories worldwide. Collectively, the data indicate that soluble HLA-G is a critical contributor to immune privilege in pregnancy and imply that this placenta-derived substance may impact other pathways leading to successful reproduction.

Evidence for expression of endogenous retroviral sequences on primate reproductive tissues and detection of cross-reactive ERVS antigens in the baboon ovary: a review.

OBJECTIVE: To review recent research findings on the specific expression of endogenous retroviral sequences (ERVS) in reproductive tissues and their possible physiological roles. ERVS have been implicated in several biological events such as induction of resistance to exogenous retrovirus invasion, involvement in placental trophoblast formation, sperm maturation and differentiation; and stimulation of local immunosuppression to protect the foetus from immunological attack. DATA SOURCES: Critical review of relevant articles and abstracts cited in international and local journals, literature searches on Medline and Medchem up to 2005. DATA SYNTHESIS: Retroviruses have been implicated in the induction of tumour and immunological disorders. Over the years, endogenous retroviruses (ERVs) and retroviral elements have been detected in the genome of many vertebrate species, including primates. The evidence for the presence of retroviruses in the primate tissues such as the placenta, ovary, breast, testis and epididymis has been documented using electron microscopic studies. Retrovirus-like particles were found budding from the basal membrane of syncytiotrophoblasts, as well as in tumour cell lines in embryonic carcinoma or teratocarcinomas. Apart from their pathological effects, recent evidence suggests that these ERVs may play useful roles in normal physiological events. RESULTS: Recent studies indicate the expression of endogenous retroviruses in the testis, epididymis, placenta and breast. However, limited data exist on the detection of ERVs in the ovary. Overall, the precise functions for ERVs in these tissues are not well understood. In the testis and epididymis, speculative functions may include among others spermatogenesis and/or sperm maturation (differentiation) whereas in placenta they are possibly associated with trophoblast fusion and locally induced immunosuppression to protect the foetus from immunological attack. Experiments in our laboratory have indicated restricted expression of retroviral antigens including baboon endogenous retroviral proteins (BERV), ERV-3, HIV-1 gp41 and HERV-K env in the baboon ovary. CONCLUSION: ERVs are specifically expressed in different mammalian reproductive tissues and may have unique physiological roles.

Evidence for expression of endogenous retroviral sequences on primate reproductive tissues and detection of cross-reactive ervs antigens in the baboon ovary: a review

Objective: To review recent research findings on the specific expression of endogenous retroviral sequences (ERVS) in reproductive tissues and their possible physiological roles. ERVS have been implicated in several biological events such as induction of resistance to exogenous retrovirus invasion; involvement in placental trophoblast formation; sperm maturation and differentiation; and stimulation of local immunosuppression to protect the foetus from immunological attack. Data sources: Critical review of relevant articles and abstracts cited in international and local journals; literature searches on Medline and Medchem up to 2005. Data synthesis: Retroviruses have been implicated in the induction of tumour and immunological disorders. Over the years; endogenous retroviruses (ERVs) and retroviral elements have been detected in the genome of many vertebrate species; including primates. The evidence for the presence of retroviruses in the primate tissues such as the placenta; ovary; breast; testis and epididymis has been documented using electron microscopic studies. Retrovirus-like particles were found budding from the basal membrane of lyncytiotrophoblasts; as well as in tumour cell lines in embryonic carcinoma or teratocarcinomas. Apart from their pathological effects; recent evidence suggests that these ERVs may play useful roles in normal physiological events. Results: Recent studies indicate the expression of endogenous retroviruses in the testis; epididymis; placenta and breast. However; limited data exist on the detection of ERVs in the ovary. Overall; the precise functions for ERVs in these tissues are not well understood. In the testis and epididymis; speculative functions may include among others spermatogenesis and/or sperm maturation (differentiation) whereas in placenta they are possibly associated with trophoblast fusion and locally induced immunosuppression to protect the foetus from immunological attack. Experiments in our laboratory have indicated restricted expression of retroviral antigens including baboon endogenous retroviral proteins (BERV); ERV-3; HIV-1 gp41 and HERV-K env in the baboon ovary. Conclusion: ERVs are specifically expressed in different mammalian reproductive tissues and may have unique physiological roles

Serological detection of adenoviruses in non-human primates maintained in a colony in Kenya.

BACKGROUND: Adenoviruses are known to cause several human diseases including acute febrile respiratory syndromes, epidemic conjunctivitis and gastroenteritis. These diseases associated with adenovirus infection affect adults and are usually more severe in infants and children. Forty-seven human adenoviruses serotypes have so far been identified adenovirus. The diversity of these viruses has delayed progress on vaccine development due to difficulties in identifying appropriate vaccine targets. To date, limited studies have been done to determine the prevalence of adenovirus infection in non-human primates with the goal of developing a non-human primate model that can be used to study the mechanisms of infection. OBJECTIVE: To determine the prevalence of enteric adenovirus infection in Kenyan non-human primates. DESIGN: A prospective study to investigate the prevalence of enteric andenovirus infection in captive non-human primates maintained in a colony. SETTING: Faecal samples were collected from monkeys trapped from different geographical areas of Kenya and also from the ones maintained in a colony at the Institute of Primate Research (IPR), Kenya. SUBJECTS: Ninety four faecal samples were collected from three species of non-human primates consisting of various ages and sex. Samples were collected from monkeys trapped from different geographical areas of Kenya and also from the ones maintained in a colony at the Institute of Primate Research (IPR), Kenya. All the faecal samples were screened for presence of adenoviruses using a commercial antigen-capture enzyme immunoassay (EIA) kit, this is an enzyme-linked immunosorbent assay (ELISA) kit designed for diagnosis of human enteric adenoviruses in stool samples. RESULTS: The highest prevalence of adenoviruses, detected by EIA kit, was in olive baboons (Papio anubis, 52.9%), followed by vervet monkeys (Cercopithecus aethiops, 48.9%) and the yellow baboons (Papio cynocephalus, 18.8%). Sub-grouping within each species (based on age and sex) indicated no significant differences (p > 0.05) in adenovirus infection signifying equal susceptibility. The prevalence of adenoviruses in vervet monkeys that were also Simian Immunodeficiency virus (SIV) seropositive was determined and shown to be 63.2%. CONCLUSION: The results of this study indicate that adenovirus infection is prevalent among non-human primates in Kenya. These findings suggest that cross species transmission in Kenyan non-human primates may be a common occurrence and there is a possibility of zoonotic transmission of adenoviruses. Furthermore, our results highlight the potential of using these non-human primates as models for testing safety and efficacy of candidate adenovirus vaccines prior to clinical trials in humans.

The olive baboon (Papio anubis): a potential animal model to study the function of human leukocyte antigen-G (HLA-G).

The human class Ib major histocompatibility complex (MHC) molecule, HLA-G, is unique in its limited polymorphism, high expression in the placenta and generation of multiple transcripts by alternative splicing. The proteins encoded by these transcripts are believed to modulate maternal-fetal immunological relationships during pregnancy. The baboon placenta expresses Paan-AG, a novel MHC molecule that is evolutionarily related to the MHC-A locus but shares unique characteristics with HLA-G. In this brief review, we present evidence suggesting that Paan-AG may be the functional homologue of HLA-G, and propose that the baboon would compromise an excellent animal model for functional studies of HLA-G proteins in human pregnancy.

The prevalence of anti-phospholipid antibodies in a selected population of Kenyan women and development of a non-human primate model.

The mechanisms by which anti-phospholipid antibodies (aPLs) may induce pregnancy losses, intrauterine growth retardation and pregnancy-induced hypertension are not clearly understood. Moreover, there is a controversy regarding the possible direct effects of these antibodies on the physiology of the placenta since the target antigens of these antibodies are intracellular antigens and are potentially inaccessible to the antibody. Also, controversy exists regarding the usefulness of the treatment regimens currently available. In this study, we present preliminary data on the prevalence of aPLs in a selected population (n = 80) of Kenyan women visiting Kenyatta National Hospital, Nairobi, Kenya for obstetrical complications including recurrent pregnancy losses. Our results showed approximately 13.8% of the patients were positive for anti-cardiolipin antibodies whereas 33.8% were positive for aPS. Additionally, we screened 72 non-human primates for presence of aPLs and our results showed that the olive baboon (Papio anubis) had the highest prevalence rate (52.2%, n = 23). Overall, our results suggest that the olive baboon may be a suitable animal model for studying the mechanism of action of the anti-phospholipid antibody and pregnancy complications associated with aPLs.

Effects of khat (Catha edulis) consumption on reproductive functions: a review.

OBJECTIVE: To review research findings on the effects of khat (Catha edulis) chewing on reproductive functions. DATA SOURCES: Retrieval and critical review of relevant articles and abstracts cited in international and local journals, literature searches on Medline and Medchem from 1961 to 2002. DATA SYNTHESIS: Analysis of published data and limited interviews of regular khat users revealed that khat chewing lowers libido in humans and may also lead to sexual impotence following long term use. In pregnant women, consumption of khat affects growth of foetus by inhibiting utero-placental blood flow and as a consequence, impairs foetal growth. CONCLUSION: Detailed studies on the effects of khat on reproduction are lacking. However, the limited available data reveal that chewing of khat has a negative impact on human reproductive health. Khat is genotoxic and has teratogenic effects on the foetus if regularly consumed by pregnant mothers. Since low birth weight is a well-established risk factor for both perinatal and young infant death, khat chewing during pregnancy may be one of the factors contributing to infant mortality in communities where khat is commonly chewed. Khat consumption affects the potency of male sexuality by affecting spermatogenesis and plasma testosterone concentration. However, the precise mechanisms by which khat may affect the male reproductive physiology have not been elucidated.

Human endogenous retrovirus-R (ERV 3) env-like antigens expressed in baboon testes and epididymides.

BACKGROUND: A substantial component of the vertebrate genome comprise of retrovirus-related sequences named as endogenous retroviruses (ERVs). The role of these ERV-related sequences in the biological processes of the host species is still unknown. However, they have been associated with tumourigenesis, autoimmune diseases and placental morphogenesis in primates. OBJECTIVE: To determine the expression of ERVs in male baboon reproductive tissues. DESIGN: The testes and other reproductive tissues from sexually immature and mature male olive baboons (Papio anubis) were investigated for the expression of endogenous retrovirus-related particles. Immunohistochemical staining was performed using antibodies raised against human immunodeficiency virus (HIV)-1/2, simian immunodeficiency virus (SIV) and human ERVs. Biochemical properties were determined by western blot, and reverse transcriptase (RTase) activity in epididymal spermatozoa, ejaculate spermatozoa and seminal fluid was evaluated. SETTING: Institute of Primate Research, Nairobi, Kenya. RESULTS: ERV3 env-like antigens were detected on spermatogenic cells in mature baboon testes and on epididymal spermatozoa. Similarly, antigens cross-reactive with antibodies to HIV structural and envelope glycoproteins were expressed in mature and juvenile baboon testes. In addition, reverse transcriptase activity was detected in ejaculate spermatozoa, seminal fluid and epididymal spermatozoa. CONCLUSION: These results indicate that retroviral-related genes were expressed in normal male baboon testes and spermatozoa, similar to humans. The functions of these ERVs in vertebrates remains unclear.

Endogenous retrovirus sequences expressed in male mammalian reproductive tissues: a review.

OBJECTIVES: To review the research findings on the expression of endogenous retroviruses and retroviral-related particles in male mammalian reproductive tissues, and to discuss their possible role in normal cellular events and association with disease conditions in male reproductive tissues. DATA SOURCES: Published findings on endogenous retrovirus (ERV) expression in vertebrate reproductive tissues. STUDY SELECTION: Relevant citations on ERVs and male reproduction by research groups worldwide. DATA EXTRACTION: Literature search on Medline and Pubmed upto the year 2000, and retrieval of relevant articles cited from international and local journals. DATA SYNTHESIS: Most of the studies demonstrated integrated retroviruses and retroviral-related sequences in human and mouse testis, epididymis and vas deferens. Endogenous retroviruses in human and mice may be associated with normal cellular differentiation and development, and carcinogenesis. In humans, one ERV family, human endogenous retrovirus-K (HERV-K) is abundantly expressed, and is associated with germ cell tumours, while ERV3 env is expressed in normal human testis. CONCLUSION: The expression of ERVs in male reproductive tissues suggests a possible role in normal and disease conditions involving the testis and epididymis. These speculative functions may include among others spermatogenesis and or sperm maturation or tumour formation. However, further studies need to be carried out to investigate specific roles of ERVs in male reproductive events.

Potential risks of viral infections in xenotransplantation.

The shortage of cadaveric human organs for transplantation may, be alleviated by the use of xenografts as a therapeutic option for end-stage organ failure. Successful attempts have been made to prevent rejection of xenograft tissues in humans. The potential spread of animal-derived pathogens to the xenograft recipient is a complication of xenotransplantation, which must be addressed. This can be complicated further by, the presence of new pathogens, new clinical syndromes, and altered behaviour of these organisms in the immunocompromised recipient. There is concern over the possible activation of latent viruses, including retroviruses, from xenograft tissues. This paper discusses the possible dangers of transmission of animal viruses to humans via xenotransplantation.

The prevalence of antibodies to simian T-cell leukaemia/lymphotropic virus (STLV-I) in non-human primate colonies in Kenya.

Retroviruses closely related to the human T-cell leukaemia/lymphotrophic virus type I (HTLV-I) have been detected in several, non-human, primate species. These retroviruses are called simian T-lymphotrophic virus type I (STLV-I). Infection with STLV-I has been associated with lymphoma and leukaemia in macaques, baboons, African green monkeys and gorillas. However, no STLV-I infection has been detected in New World primates, although STLV-II has been detected in spider monkeys. When sera from 10 species of non-human primates maintained at the Institute of Primate Research were screened for STLV-I infection, anti-STLV-I antibodies were detected in 12%, 12%, 23% and 38% of the olive baboons, yellow baboons, African green monkeys and lowland Sykes' monkeys, respectively. Western-blot studies confirmed these results. To date, no clinical disease has been linked with STLV-I infection in these colonies. The relatively high prevalence of anti-STLV-I antibodies in these non-human primates offers an opportunity for studies on the transmission, phylogenetic relationships and natural history of STLV-I in primate colonies.

Characterization of antigens expressed in normal baboon trophoblast and cross-reactive with HIV/SIV antibodies.

Electron microscopic studies have revealed the presence of endogenous retroviral (ERV) particles in normal primate placental tissues. These particles have ultrastructural similarities to type C retroviral particles and are mainly associated with the trophoblast. In normal human placental tissues, they have antigenic similarity with exogenous retroviruses, such as the human immunodeficiency virus (HIV), and may have a role to play in the regulation of cellular gene expression, syncytiotrophoblast formation or pregnancy-related immunosuppression. In this study, a panel of antibodies (polyclonal and monoclonal antibodies) against viral proteins (anti-HIV and anti-SIV) and endogenous retroviral (ERV) proteins were assessed by immunohistochemistry and immunoblotting, for their cross-reactivity with ERV particles isolated from normal baboon placental tissues. The antibodies (anti-HERV-K RT, anti-ERV3 env, anti-HIV-1 p17, anti-HIV-2 gp120) reacted positively with the syncytiotrophoblast and each antibody recognized one or two proteins of molecular weights (MW) 38, 58 or 64 kDa present in the baboon placental villous tissues and SIV-infected molt-4 Cl8 cells, but not in uninfected cells. The results of this study confirm the specific expression of retroviral cross-reactive antigens in normal baboon placental tissues and suggest placental cellular proteins may have antigenic similarity with those recognized by anti-HIV/SIV antibodies. The role of these retroviral-related proteins expressed at the maternal-fetal interface remain unclear.

Molecular characterisation of a putative endogenous retrovirus cDNA isolated from human placental tissue.

The human genome comprises of abundant DNA sequences related to endogenous retroviruses (ERV) and a variety of solitary long terminal repeats (LTRs). Substantial numbers of intact retroviral particles have been detected by electron microscopy in normal human placental villous tissue particularly in syncytiotrophoblast. Understanding the molecular structure, organisation and distribution of these ERV sequences may lead to elucidation of their possible dual function at the foetal-maternal interface; proliferation and differentiation of cytotrophoblast and induction of local pregnancy-associated immune suppression thus allowing survival of the foetal allograft. In this study, antibody probes were used to screen a human placental expression library and cDNA clones isolated were characterized by polymerase chain reaction, Southern blot hybridisation, DNA cloning and partial nucleotide sequencing. A specific 1.7kb-cDNA clone was isolated from a human placental expression library. Further characterisation showed this clone represents a single copy gene, approximately 9-10kb and did not hybridise to the env region of ERV3 human endogenous retrovirus. The 1.7kb-cDNA clone may represent a provirus co-expressed with cellular sequences.

Molecular Characterisation of a Putative Endogenous Retrovirus cDNA Isolated from Human Placental Tissue

The human genome comprises of abundant DNA sequences related to endogenous retroviruses (ERV) and a variety of solitary long terminal repeats (LTRs). Substantial numbers of intact retroviral particles have been detected by electron microscopy in normal human placental villous tissue particularly in syncytiotrophoblast. Understanding the molecular structure; organisation and distribution of these ERV sequences may lead to elucidation of their possible dual function at the foetal-maternal interface; proliferation and differentiation of cytotrophoblast and induction of local pregnancy-associated immune suppression thus allowing survival of the foetal allograft. In this study; antibody probes were used to screen a human placental expression library and cDNA clones isolated were characterized by polymerase chain reaction; Southern blot hybridisation; DNA cloning and partial nucleotide sequencing. A specific 1.7kb-cDNA clone was isolated from a human placental expression library. Further characterisation showed this clone represents a single copy gene; approximately 9-10kb and did not hybridise to the env region of ERV3 human endogenous retrovirus. The 1.7kb-cDNA clone may represent a provirus co-expressed with cellular sequences

Immunohistochemical localization of retroviral-related antigens expressed in normal baboon placental villous tissue.

Endogenous retroviral particles (ERVs) have been detected in the genome of all eukaryotes. They are generally non-pathogenic except in mice where they have been found to induce tumors and immunological disorders. The ERVs have morphological features consistent with type-C retroviral particles and are commonly expressed in normal placental villous tissues. ERVs may have a role in the regulation of placental gene expression, syncytiotrophoblast formation, or pregnancy-related immunosuppression. In this study, well-characterized antibodies (monoclonal and polyclonal antibodies) raised against retroviral proteins (anti-HIV and anti-SIV) and endogenous retroviral (ERV) particles were assessed for their cross-reactivity (by using immunohistochemistry) with normal baboon placental and other adult tissues. The monoclonal antibodies to exogenous retroviral proteins (anti-HIV-2 gp120, anti-HIV-1 gp41, anti-SIVmac p27, anti-HIV-1 RT, and anti-HIV-2 core protein) showed specific immunohistochemical reactivity with the syncytiotrophoblast. Antibodies to endogenous retroviral gene products (anti-ERV3 env, anti-HERV-K RT, and anti-HERV-K env) also reacted in a similar manner and did not cross-react with other adult tissues. These studies have shown that retroviral-cross-reactive proteins are expressed in baboon placental syncytiotrophoblast and may have a role to play at the feto-maternal interface.

Expression of endogenous retroviral genes in germ cells and embryonic tissues in primates.

The presence of endogenous retroviral sequences (ERVs) in vertebrate genomes is well documented. ERVs have been found to be generally inactive and non-infective except in certain pathological conditions. However, baboon endogenous rectrovirus (BaEV) has been shown to be infective in vitro, and ERV particles have been isolated from normal placental villous tissues of most primates. These particles have ultra-structural features similar to type C retroviruses and are cross-reactive with antibodies to exogenous retroviruses such as HIV. Their function is not yet known. This review examines their possible role in modulation of the immune system during pregnancy and in the syncytiotrophoblast in primates.

Characterisation of retroviral-related antigens expressed in normal baboon placental tissues.

The genomes of all eukaryotes contain multiple copies of DNA sequences that are related to sequences found in infectious retroviruses. Endogenous retroviruses (ERVs) are generally non-pathogenic although they have been implicated in the induction of tumours and immunological disorders. The ERVs have morphological features consistent with type-C retroviral particles and are expressed in normal placental tissue in most mammals. They have antigenic similarity with exogenous retroviruses such as HIV-1 and may have a role to play in the regulation of cellular gene expression, syncytiotrophoblast formation or pregnancy-related immunosuppression. Some of the human endogenous retroviruses have been well-characterised. Among the non-human primates, the baboon endogenous virus (BaEV) is the only endogenous retrovirus so far which has been shown to be effective in vitro. The entire nucleotide sequence of BaEV has been determined. It has been shown to have a chimeric genomic structure of about 8 kb long. BaEV particle expression in placental tissues has been demonstrated using electron microscopy. However, to date, very little work has been done to evaluate the expression of retroviral-related antigens in normal baboon tissues. In this study, mouse polyclonal antibodies were produced against isolated baboon placental ERV particles and characterised using immunohistochemistry and immunoblotting techniques. Most of the anti-BERV antibodies displayed specific immunoperoxidase staining on placental syncytiotrophoblast and cross-reacted with exogenous retroviral proteins on immunoblot analyses. Reverse transcriptase (RTase) activity associated with sucrose gradient-purified placental retroviral-like particles were also demonstrated. These studies indicate that endogenous retroviral particles are expressed in baboon placental villous tissue and suggest retroviral proteins may play an immunomodulatory role at the maternal-foetal interface.