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J Menezes

Publications and source records attributed to J Menezes.

At least 19 recordsLinked to original sources

New global defect structures.

We investigate the presence of defects in systems described by real scalar field in (D,1) spacetime dimensions. We show that when the potential assumes specific form, there are models which support stable global defects for D arbitrary. We also show how to find first-order differential equations that solve the equations of motion, and how to solve models in D dimensions via soluble problems in D=1. We illustrate the procedure examining specific models and finding explicit solutions.

Journal Article↗

Virus load correlates inversely with the expression of cytotoxic T lymphocyte activation markers in HIV-1-infected/AIDS patients showing MHC-unrestricted CTL-mediated lysis.

Cytotoxic T lymphocytes (CTL) are key players to suppress viral load (VL) but CTL responses become compromised with progression of HIV-infection/AIDS. Some progressors develop MHC-unrestricted CTL with anti-CD4+ cytocidal activity. Immune activation status of these CTL and its significance in disease progression are unknown. To determine the relationship between VL and T cell activation, a cross-sectional study was carried out using blood samples from 13 HIV-1-infected/AIDS patients at various stages of progression and seven age-matched seronegative controls. We examined expression of HLA-DR and CD38 activation markers on purified CTL. MHC-unrestricted killing by these CTL was also evaluated against uninfected, allogeneic CD4+ T cells as well as several human cell lines. The expression of activation markers correlated inversely (rs = - 0.91, P < 0.0001) with VL of the subjects. CTL effectors of these patients killed targets expressing or lacking CD4+, independently of MHC class I recognition. Interestingly, the patients with higher VL showed an increased number of gammadeltaTCR-bearing CTL in blood and their MHC-unrestricted killing activity was blocked significantly (P < 0.01) by gammadeltaTCR-specific monoclonal antibody. CD3+ T counts of these patients were also consistently subnormal. Inverse correlation between VL and CD8+ T cell activation markers seems to be an indicator of CTL-associated immunopathogenesis in HIV patients with elevated gammadeltaCTL in the peripheral blood.

Acquired Immunodeficiency Syndrome↗

Interleukin 15-mediated induction of cytotoxic effector cells capable of eliminating Epstein-Barr virus-transformed/immortalized lymphocytes in culture.

BACKGROUND: Interleukin 15 (IL-15) activates cytotoxic lymphocytes and drives the expansion of memory T cells. Its role in immune control of virus-transformed cells and other tumor cells remains to be elucidated. We investigated the role of IL-15 in controlling Epstein-Barr virus (EBV)-transformed/immortalized lymphocytes in culture. EBV is a highly potent lymphocyte-transforming and opportunistic oncogenic herpesvirus associated with several human tumors. METHODS: Peripheral blood mononuclear cells (PBMCs) from healthy donors were infected with EBV and cultured with either IL-15 or IL-15 plus anti-IL-15 antibodies for 3-4 weeks. We monitored EBV-induced transformation by assessing the clearly visible cell clusters by microscopy and analyzing the expression of EBV-encoded latent membrane oncoprotein-1 (LMP-1) and the EBV nuclear antigen (EBNA) complex by immunoblotting and immunofluorescence techniques, respectively. We depleted EBV-infected cultures of PBMCs of specific effector cell populations to investigate the effector cells involved in mediating IL-15 effect. RESULTS: The presence of IL-15 resulted in the complete elimination of EBV-transformed cells in PBMC cultures. Western blot and immunofluorescence analyses performed 3-4 weeks after infection showed no detectable levels of LMP-1 and EBNA in IL-15-treated EBV-infected cultures, whereas IL-15-untreated EBV-infected cultures and IL-15/anti-IL-15-treated cultures expressed both proteins. IL-15 mediated its anti-EBV effect through early and late response mechanisms, i.e., by first activating natural killer (NK) cells and subsequently inducing cytolytic NK-T cells. The presence of anti-IL-15 neutralizing antibodies abrogated IL-15's effect on both mechanisms. CONCLUSION: In vitro, IL-15 mediated complete elimination of EBV-infected/transformed lymphocytes via successive activation of NK and NK-T cytotoxic effectors. If these in vitro findings reflect in vivo mechanisms, then IL-15 might be considered for cytokine-based immunotherapy in patients with EBV-associated lymphoproliferative disorders/malignancies.

B-Lymphocytes↗

Epstein-Barr Virus and its glycoprotein-350 upregulate IL-6 in human B-lymphocytes via CD21, involving activation of NF-kappaB and different signaling pathways.

Epstein-Barr virus (EBV) is a ubiquitous and highly immunotropic gamma herpesvirus that infects more than 90 % of humans worldwide. Its pathogenicity leads to a number of diseases including tumors that result from EBV's ability to readily transform B-lymphocytes and, to a lesser extent, epithelial cells. EBV utilizes CD21/CR2 as its receptor on B cells to initiate the infection process. EBV binds to CR2 through its major envelope glycoprotein-350 (gp350) and is also a remarkable immunomodulating agent. We had previously shown that EBV is capable of modulating the synthesis of a number of cytokines. We now show that while both purified recombinant gp350 (rgp350) and EBV upregulate IL-6 mRNA synthesis in B cells, EBV-induced IL-6 gene activation occurs for a significantly longer period of time (i.e. 12 hours for EBV as compared to 6 hours for rgp350). Moreover, the half-life of EBV-induced IL-6 mRNA was also significantly longer (10 hours) than that of mRNA induced by rgp350 (about 6 hours). Both EBV and gp350 enhance the binding of the NF-kappaB transcription factor, as determined by band-shift and augment NF-kappaB-mediated activation of a CAT reporter plasmid. Furthermore, we demonstrate that while the activation of IL-6 gene expression by gp350 is mediated primarily by the protein kinase C pathway, EBV can mediate its effects through multiple signaling pathways. To our knowledge this is the first report showing that the binding of a herpesvirus envelope glycoprotein to CR2 on human B cells results in the activation of the NF-kappaB transcription factor leading to the upregulation of IL-6 gene expression in these lymphocytes.

B-Lymphocytes↗

Evidence for a correlation between antibody-dependent cellular cytotoxicity-mediating anti-HIV-1 antibodies and prognostic predictors of HIV infection.

Using our gp120/41-expressing, NK cell activity-resistant CEM.NKR cell clones as targets in HIV-1-specific antibody-dependent cellular cytotoxicity (ADCC) assays, we demonstrate here that the serum titers of anti-HIV-1 ADCC antibodies bear a significant (P < 0.05) positive correlation with the peripheral blood CD4+ T cell counts and a negative one with the number of copies of HIV-1 RNA in the plasma of HIV-infected individuals. These findings underscore the importance of these antibodies as a protective immune parameter in these infections.

Antibody-Dependent Cell Cytotoxicity↗

Activated human platelets express Fas-L and induce apoptosis in Fas-positive tumor cells.

In addition to their role in hemostasis and thrombosis, platelets are important modulators of immune and inflammatory responses. We provide evidence here that human platelets contain abundant quantities of Fas-L, and upon activation, they express it on their surface as well as release it into medium. This surface-expressed Fas-L is biologically active and can induce apoptosis in Fas-positive human tumor cells. Therefore, activated platelets may represent an important player in Fas/Fas-L-mediated apoptosis.

Apoptosis↗

Modulation of expression of the MHC class I-binding natural killer cell receptors, and NK activity in relation to viral load in HIV-infected/AIDS patients.

The natural killer (NK) cells play an important role in viral infections via their spontaneous cytolytic activity against virus-infected cells as well as via secreting a variety of soluble mediators. The MHC class I-binding NK receptors of these cells have emerged as the most important regulators of the effector activities of cytolytic cells (both NK and CTL). We have studied the modulation of NK activity and the expression of NK receptors in HIV-infected/AIDS patients and report here that the NK activities of the patients with the lowest plasma HIV load were minimal and vice versa, suggesting a decrease in this activity following suppression of HIV replication. Interestingly, the NK activity correlated negatively with the peripheral blood CD4+ T-cell counts of these patients. Furthermore, these patients showed decreased percentages of CD56+ cells expressing NK receptors of the immunoglobulin superfamily, whereas the percentages of CD8+ cells expressing these receptors were increased. Moreover, the expression of C-type lectin-like NK receptor-associated invariant molecule CD94 was increased on CD8+ cells in these patients as compared with HIV-seronegative controls. These changes in the expression of NK receptors were also evident within groups of these patients having different viral loads. These results show, for the first time, decreased innate immunity and changes in the expression of NK receptors on cytolytic cells in relation to viral burden in HIV-infected/AIDS patient.

Adult↗

Characterization of human gamete centrosomes for assisted reproduction.

Centrosomes of human gametes need to be characterised in more detail, since it was recently shown in 1991 that the human embryo inherits the dominant paternal centrosome at fertilization. Thus there has been a resurgence of centrosomal research in the last decade. The sperm centrosome, containing a single centriole, is preserved and dominant while the egg centrosome is reduced and inactivated during oogenesis, since there should be only one functional centrosome to ensure normal development. This presentation reviews the ultrastructure of gamete centrosomes and attempts to define their respective roles in assisted fertilization. Human testicular and ovarian tissues, sperm, eggs and zygotes were examined by routine TEM. Developing sperm cells and oogonia have two functional centrioles (diplosomes) in their centrosomes, showing the typical "9+0" organisation of microtubule triplets--common to somatic cells. The diplosomes are surrounded by pericentriolar material (PCM), which nucleate microtubules (MT) that organize the cytoskeleton and mitotic spindles. During spermiogenesis, when the spermatids transform into mature sperm, there is a partial reduction of the male centrosome, during which the proximal centriole (PC) is retained intact in the sperm neck, proximal to the nucleus, while the distal centriole (DC) which gave rise to the sperm flagellum, is partially reduced and merges distally with the sperm axoneme in the midpiece and tail. The proximal functional centriole is located in a "black box" in the neck, composed of the capitulum beneath the basal plate and flanked laterally by 9 segmented columns and shows the typical "9+0" organisation of triplets of MT. The PC contains dense material both within and outside the triplets, and the central doublet of axonemal MT terminates in a clump of dense material below its lower vault. The DC is disorganised proximally showing loss of triplets and cannot function as a typical centriole, since the central doublets of axonemal MT traverse through it. Oogonia present a pair of well-defined centrioles, which are involved in cell division. These are lost during oogenesis and the mature oocyte is devoid of centrioles, as in most mammals. Neither does the human oocyte have granular centrosomal material at meiotic spindle poles, in contrast to mouse oocytes which have a dominant maternal centrosome. Thus the oocyte centrosome is greatly reduced and inactivated. Functional centrosomal structure is, however, restored after fertilization in the zygote with some maternal input around the sperm centriole, which duplicates at the pronuclear stage, forms a sperm aster and proceeds to form the first mitotic spindle. This is the ancestor of centrosomes in embryonic, foetal and adult somatic cells. In 1991 (Sathananthan, 1991), we postulated that sperm centrosomal dysfunction could lead to aberrant embryonic development based on centriolar defects in sperm with impaired motility. This hypothesis has now gained acceptance and further evidence to support this theory of infertility is presented.

Centrosome↗

Thrombin induces apoptosis in human tumor cells.

Thrombin is a serine protease that is produced during the coagulation process and plays an essential role for hemostasis, thrombosis and wound healing. It is a potent activator of platelets, induces proliferation of a wide variety of normal and malignant human cells, and enhances their invasiveness and metastatic potential. We studied the effect of thrombin on the proliferation of a wide variety of human tumor cells and report here that, at low concentrations, thrombin induces proliferation of these cells. However, at higher concentrations, thrombin inhibited their proliferation. We show that this inhibition of cell proliferation was due to apoptosis of the tumor cells. The thrombin-mediated apoptosis was inhibited significantly by its specific inhibitor, hirudin. Furthermore, no consistent pattern of induction and/or modulation of p53, p21 and bcl-2 was observed in the thrombin-mediated apoptosis. To our knowledge, this is the first report to describe the pro-apoptotic effects of thrombin on human tumor cells and may have implications for chemotherapy in cancer patients and for the pathogenesis of AIDS as well.

Antineoplastic Agents↗

Binding of the Epstein-Barr virus major envelope glycoprotein gp350 results in the upregulation of the TNF-alpha gene expression in monocytic cells via NF-kappaB involving PKC, PI3-K and tyrosine kinases.

Epstein-Barr virus (EBV) is a human herpesvirus that interacts with various immunocompetent cells that carry the EBV receptor (CD21/CR2). EBV binds to CR2 through its major envelope glycoprotein 350 (gp350). Previously we had demonstrated that EBV and other human herpesviruses are capable of modulating cytokine synthesis through the deregulated expression of cytokine genes interleukin-1 (IL-1), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), and interleukin-2 (IL-2). Here we show that, in contrast to infectious EBV, purified recombinant gp350 upregulates TNF-alpha gene expression in human monocyte/macrophages (M/M) as well as in a monocytoid cell line, U937. Our results also demonstrate that this increased expression is due to both enhanced transcription and stability of TNF-alpha mRNA in gp350-treated cells. The specificity of this effect is evidenced by the fact that pre-incubation of cells with anti-CR2 monoclonal antibody OKB7, which blocks binding of gp350 to CR2, inhibits the above mentioned effects of gp350. Furthermore, we demonstrate that activation of TNF-alpha by gp350 is mediated by NF-kappaB through signal transduction pathways involving PKC, PI3-K and tyrosine kinases. To our knowledge this is the first report describing the modulation of TNF-alpha gene expression by the EBV-gp350 molecule following its interaction with the viral receptor CR2 on cells of the monocytic lineage.

Antibodies, Monoclonal↗

Analysis and significance of anti-latent membrane protein-1 antibodies in the sera of patients with EBV-associated diseases.

Anti-latent membrane protein-1 (LMP-1) is an EBV-encoded type III integral membrane protein with oncogenic potential that is expressed most consistently in various EBV-associated malignancies. Unlike many other EBV proteins, LMP-1 Abs have rarely been demonstrated in EBV-associated disease conditions. We established a high level LMP-1-expressing cell clone and used it for the detection, quantitation, and characterization of these Abs in various human sera in immunoblots and ELISA. Our results demonstrate that, in contrast to the commonly held notion, LMP-1 induces significant humoral immune responses in EBV-associated malignant conditions especially in nasopharyngeal carcinoma (NPC) patients in whom >70% sera are positive for these Abs, and their titers correlate with the clinical condition of the tumors. Interestingly, anti-LMP-1 Abs of IgA isotype were found only in NPC patients. These Abs were absent from the sera of infectious mononucleosis and chronic EBV infection patients, whereas a small fraction ( approximately 5%) of the healthy, EBV-seropositive individuals were positive for them; however, their OD values were much lower than those of NPC patients. These studies demonstrate, for the first time, the potential significance of LMP-1-specific Abs for the diagnosis and prognosis of EBV-associated malignancies, especially of NPC.

Antibodies, Viral↗

Innate immune response of the human host to exposure with herpes simplex virus type 1: in vitro control of the virus infection by enhanced natural killer activity via interleukin-15 induction.

Infections with herpes simplex virus type 1 (HSV-1) in humans and in animal models are accompanied by enhanced natural killer (NK) activity. In vitro, HSV-1 also enhances the NK activity of human peripheral blood mononuclear cells (PBMC). The molecular basis of this enhanced NK activity, however, is not well characterized. We investigated the role of human interleukin-15 (IL-15) in this phenomenon and report here that HSV-1-mediated enhanced NK activity was abrogated by neutralizing antibodies for IL-15 but not for other cytokines (i.e., IL-2, IL-12, gamma interferon [IFN-gamma], tumor necrosis factor alpha, or IFN-alpha). Anti-CD122 antibodies which block signaling through IL-2 receptor beta chain, and therefore neutralize the effects of IL-15 (and IL-2), also abrogated this enhancement. Furthermore, HSV-1 increased the levels of IL-15 mRNA and the production of IL-15 in HSV-1-infected PBMC cultures. The neutralization of IL-15 in cocultures of PBMC with HSV-1-infected cells significantly increased HSV-1 production. These results strongly suggest a role for IL-15 in the HSV-1-mediated in vitro enhancement of NK activity and in the PBMC-mediated suppression of HSV-1 replication.

Antibodies, Monoclonal↗

Elevated serum transforming growth factor beta1 levels in Epstein-Barr virus-associated diseases and their correlation with virus-specific immunoglobulin A (IgA) and IgM.

Transforming growth factor beta (TGF-beta) is an immunosuppressive cytokine which can induce immunoglobulin A (IgA) switch and Epstein-Barr virus (EBV) replication in latently infected cells. Here we report elevated serum levels of TGF-beta in various EBV-associated diseases correlating positively with EBV-specific IgA titers and negatively with IgM titers, suggesting a role for this cytokine in the pathogenesis of these diseases.

Antibodies, Viral↗

Up-regulation of NK cytotoxic activity via IL-15 induction by different viruses: a comparative study.

IL-15 is a recently identified cytokine that belongs to the four alpha-helix bundle cytokine family and possesses biological activities similar to those of IL-2. Its ability to induce effectors of NK activity suggests its involvement in innate immunity. In this study, we analyzed the effect of different viruses (HSV, EBV, respiratory syncitial virus, vesicular stomatitis virus, influenza virus, reovirus, and Sendai virus) on the up-regulation of NK activity in vitro. Exposure of human PBMC to the these viruses resulted in an immediate up-regulation of NK activity of PBMC via IL-15 induction; this effect was abrogated in the presence of mAbs to IL-15. Results of experiments conducted in parallel using mAbs to IL-15, as well as to other cytokines (IL-2, IL-12, IFN-gamma, and TNF-alpha), clearly indicated that IL-15 was specifically responsible for the observed effect. Furthermore, supernatants of virus-infected PBMC cultures significantly enhanced NK activity of uninfected PBMC in vitro. An increase of IL-15 protein levels 20 h postinfection was also confirmed in a bioassay using the IL-2-dependent cell line CTLL. Kinetic analysis of IL-15 mRNA expression using a semiquantitative RT-PCR revealed that the level of IL-15 messages peaked at different time points (up to 12 h) postinfection, depending on the nature of the virus. Taken together, these results suggest that the IL-15 response of the host to viral infection and the subsequent NK cell activation represent an important effector mechanism of the innate immune surveillance of the host against viral infections.

Cell Line↗

[In vitro cytokine induction in leukocytes by human herpesvirus 7].

Virus infections of the immune system may alter the normal cytokine profile, which leads to symptoms of illnesses. HHV-7 revealed lately has been known as one of the causative agents of exanthem subitum and pityriasis rosea, while its latent infection can be reactivated in immunocompromised conditions. Their pathomechanism has not been explored yet, therefore, the production of some cytokines by separated leukocytes was studied upon primary and secondary infections. After combined treatments with live or inactivated viruses and/or mitogens, the cytokines were quantitated in the supernatant of cells by sandwich ELISA. It was established that, individual cytokines were produced at maximal output at different times postinfection. The quantity of their molecules depended on the primary or repeated infections. Inactivated viral particles also induced cytokine release. Production of IL-2 and IFN-gamma due to mitogen activation could be augmented in primary, but was diminished in secondary infections. Release of TNF-alpha and IL-1 beta was parallel, but the combinatory effect with mitogens increased the quantity of IL-1 beta. HHV-7 induced early IL-10 production, which is known to inhibit cytokine release from helper lymphocytes and consequently might play a role in those acute inflammatory skin diseases mentioned above. In contrast to the related HHV-6 found in severe immunosuppressed conditions, the effect of HHV-7 on the balanced production of different cytokines seems to be related to the mild symptoms of skin disorders caused and that of moderate immunosuppression upon reactivation.

Cytokines↗

Elevated serum levels of transforming growth factor beta1 in Epstein-Barr virus-associated nasopharyngeal carcinoma patients.

Nasopharyngeal carcinomas (NPCs) of non-keratinizing type are strongly associated with Epstein-Barr virus (EBV). EBV and its gene products induce the synthesis and/or release of transforming growth factor beta1 (TGF-beta1) from human cells and platelets. TGF-beta1 is an immunosuppressive cytokine, and many tumors are known to secrete it, to counter the host immune response. To determine the potential role of this cytokine in the pathogenesis of NPC, 53 serum samples from patients with EBV-associated NPC and 20 from healthy donors were analyzed for total and active TGF-beta content using ELISA. Serum samples for TGF-beta content were also analyzed from NPC patients at different clinical stages of the tumors. Significantly higher (p < 0.01) levels of active and total TGF-beta were found in the sera of NPC patients than in control sera. The ratio of active:total TGF-beta was also significantly (p < 0.01) increased in the NPC sera. Levels of this cytokine were also significantly higher (p < 0.05) in the sera of patients with advanced stages of tumor compared to patients with earlier stages. Furthermore, higher levels were seen in patients with relapsing than with remitting tumors; even higher levels were observed in NPC patients who died of the tumor. Our data suggest a role of this cytokine in the pathogenesis of NPC; therefore, it may prove to be a valuable biomarker molecule for the diagnosis and prognosis of NPC. Int. J. Cancer (Pred. Oncol.) 84:396-399, 1999.

Biomarkers, Tumor↗