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C D Pauza

Publications and source records attributed to C D Pauza.

At least 55 records · Page 3Linked to original sources
PubMed1996

Pertussis toxin induces lymphocytosis in rhesus macaques.

Lymph nodes and other solid tissues of the immune system are the principal sites for antigen presentation and lymphocyte activation. Lymphocytes in peripheral blood recognize the high endothelial venules within lymphoid tissues and cross from blood to tissue by the process of extravasation. Pertussis toxin is known to block extravasation and cause lymphocytosis in murine models but has not been studied extensively in nonhuman primates. We used intravenous injection of soluble pertussis toxin to induce a transient lymphocytosis in rhesus monkeys. The increase in total white blood cells was proportionally greater for lymphocytes than for polymorphonuclear cells and the CD4+ lymphocyte subpopulation increased more than the CD8+ cell population. The presence of immature polymorphonuclear cells suggested some activation of bone marrow. Clinical chemistry studies revealed an effect of pertussis toxin on liver function. Pertussis toxin is a powerful immunomodulatory agent that can disrupt and reorganize solid lymphoid tissues.

Animals↗
PubMed1996

Tracking of dye-labeled lymphocytes in rhesus macaques.

Lymphocytes were isolated from rhesus monkeys and marked with a fluorescent lipophilic dye to monitor their distribution in vivo. Dye-labeled cells were either monitored by blood draws over a three-month period, or identified within peripheral organs upon autopsy. Lymphocyte labeling conditions were optimized. Dye-labeled lymphocytes could be detected in the circulation for at least 100 days by flow cytometry and fluorescence microscopy. Activated lymphocytes were removed from the circulation more rapidly than lymphocytes that had not been activated.

Animals↗
PubMed1996

Intrarectal transmission of simian immunodeficiency virus in rhesus macaques: selective amplification and host responses to transient or persistent viremia.

Intrarectal simian immunodeficiency virus (SIV) infection in rhesus macaques is a model for sexual transmission of primate retroviruses. Phylogenetic studies on envelope gene sequences that were present in blood following intrarectal SIV inoculation provided evidence for selective amplification of a subset of viruses present in the inoculum and defined one amino acid sequence uniquely associated with intrarectal infection. Both persistent and transient viremia states were observed after intrarectal infection. Immune responses in persistently infected animals accounted for slower rates of disease progression despite the presence of highly pathogenic viruses that were documented by transfusion studies. Transient viremia elicited protective immunity against subsequent intrarectal virus challenge but did not protect against intravenous virus challenge. Transient viremia usually but not always led to self-limiting infection. In one animal, we documented a relapse to active viremia long after the initial transient viremia. SIV transmission across mucosal barriers affects pathogenesis in the short term by limiting the types of viruses established in the host and in the longer term by establishing host responses that slow disease progression despite the presence of highly pathogenic viruses in blood.

Amino Acid Sequence↗
PubMed1995

Therapeutic approaches to HIV infection based on virus structure and the host pathogen interaction.

The HIV-1 infection of central nervous system, with attendant neuropathy and dementia, poses a unique challenge for antiviral therapy. For practical considerations, it is important to define carefully the precise therapeutic objectives. (1) Is it necessary to inhibit spreading HIV-1 infection in the central nervous system? (2) What is the role of inflammatory responses in central nervous system disease during HIV-1 infection? (3) Is there a correlation between pathology and dementia? (4) Are virions or virus gene products toxic in the central nervous system? (5) Is there a role for immune suppression and opportunistic pathogens in AIDS dementia? The development of therapeutic agents for HIV-1 infection is guided by our knowledge of virus structure, the function of viral proteins, the interactions with host components, and detailed features of the virus life cycle. In each case, unique features of the virus can be identified and established as targets for unique antiviral compounds. Drugs acting as inhibitors of virus enzymatic functions are plagued by the rapid development in vivo of drug-resistant virus variants, although combination or alternating chemotherapeutic regimens may obviate some of these concerns. Novel approaches to inhibiting virus are flourishing. In vitro studies show the value of agents as diverse as molecular decoys for tat activity to efforts to mutagenize integrated proviruses by modified oligonucleotides that form triple helices with chromosomal genes. As each particular clinical situation is better defined, the design and application of these agents can be refined to inhibit HIV-1 replication and reduce the associated morbidity.

Antiviral Agents↗
PubMed1995

Gamma delta T cells in rhesus monkeys and their response to simian immunodeficiency virus (SIV) infection.

Recent reports of the increase in peripheral blood gamma delta T cells in HIV+ patients prompted us to examine the gamma delta T cell system in rhesus monkeys (Macaca mulatta) and the responses of these cells to SIV infection. Our results reveal differences in the gamma delta T cell subset composition and their expression of CD8 in the peripheral blood of monkeys and humans. The outgrowth of simian gamma delta T cells in response to Daudi cells is similar to that in humans, but the exposure to IL-2 stimulates preferentially the simian V delta 1 subset rather than the V gamma 9/V delta 2 subset as found in humans. Upon SIV infection of the monkeys, we observed a transient increase of the percentage of total gamma delta T cell and the V gamma 9 subset. gamma delta T cells from infected animals also express more activation markers such as CD69, CD44 and the memory marker CD45RO. However, they respond to a lesser degree to Daudi or IL-2 stimulation in the outgrowth experiments compared with uninfected animals, although the subset composition of total gamma delta T cells is similar in infected and uninfected animals. The results clearly indicate that gamma delta T cells in rhesus monkeys are influenced by SIV infection. The detailed analysis of the gamma delta T cell response to SIV infection can serve as a model for understanding human gamma delta T cell responses to HIV infections.

Animals↗
PubMed1994

2-LTR circular viral DNA as a marker for human immunodeficiency virus type 1 infection in vivo.

The unintegrated viral DNA found in human immunodeficiency virus type 1 infection includes linear and circular forms. We targeted the circular form containing two copies of the viral long terminal repeat (2-LTR circle) and developed specific assays to detect this molecule in peripheral blood mononuclear cells from HIV-infected patients. In vitro HIV-1 infection of peripheral blood mononuclear cells showed rapid accumulation and rapid decay of 2-LTR circular viral DNA. Examination of 2-LTR circular viral DNA levels provides a view of spreading infection based on a viral DNA form that is structurally distinct and has a known, short half-life in infected cells. In patients not receiving antiviral therapy, the levels of 2-LTR circular viral DNA and total viral DNA were significantly correlated to CD4 cell counts. Similar correlations were not observed in patients receiving zidovudine (AZT), didanosine (ddA), or zalcitabine (ddC).

Base Sequence↗
PubMed1994

Selective amplification of simian immunodeficiency virus genotypes after intrarectal inoculation of rhesus monkeys.

Animal models for sexual transmission of human immunodeficiency virus can define the influences of virus type, dose, and route of inoculation on infection and clinical outcome. We used an uncloned simian immunodeficiency virus stock (SIVmac) to inoculate cells in vitro and to inoculate rhesus monkeys by intravenous and intrarectal routes. The distribution of virus genotypes present in each of these infection examples was characterized by DNA sequence analysis of viral long terminal repeats (LTRs). Our analysis of LTR sequences from in vitro and in vivo infections revealed three main genotypes: one genotype was observed only for in vitro infection, and two other genotypes were recovered only from infected animals. By comparing animals inoculated with high intrarectal doses of SIVmac and those inoculated with low doses, we demonstrated that unique subsets of the stock were selected after intrarectal infection. Our findings indicate that minor genotypes present in the stock cross the rectal mucosa and are amplified selectively to become prominent in peripheral blood mononuclear cells from acutely infected animals. Studies with a molecular recombinant of SIV and human immunodeficiency virus type 1 sequences, SHIV, showed that viral LTR sequences do not undergo especially rapid sequence variation or rearrangement after intrarectal inoculation. The mucosal barrier exerts a significant influence on infection and disease progression by reducing the efficiency of SIVmac infection and by permitting distinct, pathogenic genotypes to become established in the host.

Animals↗
PubMed1993

Human immunodeficiency virus infection of human brain capillary endothelial cells occurs via a CD4/galactosylceramide-independent mechanism.

Neuropathologic studies of AIDS patients have shown that brain capillary endothelial cells are a cellular target for human immunodeficiency virus (HIV) in vivo. We have established in vitro cultures of primary human brain capillary endothelial (HBCE) cells. Using this model system, we have shown a significant HIV infection of HBCE cells that is productive yet noncytopathic. The infection is mediated by a cellular interaction with gp120 that does not involve CD4 or galactosylceramide. HIV infection of HBCE cells may contribute to AIDS-associated neuropathology by disturbing the physiology of the endothelium and directly or indirectly facilitating dissemination of virus to the central nervous system.

CD4 Antigens↗
PubMed1993

Derivatives of 4-amino-3,6-disulfonato-1,8-naphthalimide inhibit reverse transcriptase and suppress human and feline immunodeficiency virus expression in cultured cells.

We have developed a series of 4-amino-3,6-disulfonato-1,8-naphthalimide (ADSN) derivatives in an attempt to create nontoxic compounds effective against lentivirus infections. The ADSN derivative Lucifer Yellow CH ([N-(hydra zinocarbonyl)amino]-4-amino-3,6-disulfonato-1,8-naphthalimid e) (LYCH) was chosen as a parent compound because of its low toxicity in vivo and in vitro and its tendency to accumulate in monocyte/macrophages, a major reservoir for lentiviruses in vivo. Several ADSN derivatives inhibited reverse transcriptases (RTs) from human immunodeficiency virus type 1 (HIV-1) and feline immunodeficiency virus (FIV). Viral expression in HIV-infected human peripheral blood mononuclear cells was inhibited by noncytotoxic concentrations of two ADSN derivatives, designated A4 (biphenyl-4,4'-dicarboxaldehyde, Lucifer Yellow CH monohydrazone; EC50 = 29 microM after 6 days) and H4 (biphenyl-4,4'-dicarboxaldehyde, Lucifer Yellow CH dihydrazone; EC50 = 5.61 microM). A4 effectively suppressed the expression of FIV in infected Crandall feline kidney fibroblasts (CRFK) at 46.2 microM, reducing the RT levels by 97% after 19 days under conditions allowing direct cell-to-cell transmission of the virus. The viability of drug-treated FIV-infected CRFK cells increased significantly in the presence of A4 relative to the viability of untreated virus-infected cells. In contrast to A4 and H4, LYCH (which lacks the appended aromatic rings characteristic of A4 and H4) had no inhibitory effects on either virus and did not inhibit RT ex vivo. However, flow cytometry studies showed that both A4 and LYCH accumulate in two cell types that can support lentiviral infections: U937 human monocytic leukemic cells that have been induced to differentiate by using tetradecanoyl phorbol acetate, and CRFK cells.

1-Naphthylamine↗
PubMed1993

Vitamin D3 compounds regulate human immunodeficiency virus type 1 replication in U937 monoblastoid cells and in monocyte-derived macrophages.

We studied the effects of vitamin D3 compounds on the replication of human immunodeficiency virus type 1 (HIV-1) in the monoblastoid cell line U937 and in primary monocyte-derived macrophage cultures to understand how modulators of monocyte/macrophage effector function might affect the pathogenesis of HIV-1 infection. U937 cell cultures exposed to 1, alpha 25-dihydroxyvitamin D3 prior to HIV-1 infection showed enhanced virus replication that was apparently due to increased cellular resistance to viral cytopathic effects; a marked inhibition of virus replication was noted in cells exposed to 1 alpha,25-dihydroxyvitamin D3 subsequent to infection. Exposure of blood-derived monocyte/macrophages to vitamin D3 compounds prior to infection also affected virus growth; in most cases, substantial inhibition of HIV-1 replication was noted in vitamin D3-treated macrophage cultures. Our results demonstrate that vitamin D3 compounds with recognized abilities to induce cellular differentiation can modulate HIV-1 infection of human macrophages.

Calcifediol↗
PubMed1992

Extrachromosomal human immunodeficiency virus type 1 sequences are methylated in latently infected U937 cells.

Long-term human immunodeficiency virus type 1 (HIV-1) infection of the human monocytic cell line U937 resulted in a progressive loss of infectivity that was correlated with the accumulation of stable, extrachromosomal forms of viral DNA. Viral latency was also characterized by reduced levels of HIV-1 transcription. The structure and activity of extrachromosomal viral DNA (E-DNA) in a fully latent U937 cell line was investigated by molecular cloning and DNA transfection. The resulting 18-kb E-DNA clone was composed of an intact HIV-1 sequence flanked by 7 kb of host sequence to one side and 1 kb of host DNA to the other side. This configuration is the result of retroviral integration into a highly repetitive element of the Alu family. Transfection of the E-DNA clone resulted in the production of infectious virus, indicating that viral latency was not the result of mutations in the HIV-1 genome. Analysis of CCGG sites revealed extensive de novo methylation of viral sequences present within E-DNA. These results suggest that modification of extrachromosomal viral DNA sequences is a mechanism for HIV-1 latency in long-term infected U937 cells.

Base Sequence↗
PubMed1991

The endocytic pathway for human immunodeficiency virus infection.

We have presented data supporting the view that HIV infection of T cells and monocytes proceeds via receptor-mediated endocytosis. Biochemical analysis of the pathway for internalization of 32P-labeled HIV revealed the presence of intact, intercellular virions and provided evidence for subsequent uncoating of these particles and release of viral RNA to the cytoplasm. Electron microscopy demonstrated virions within vesicles and documented fusion between viral and endosomal membranes. Others have provided evidence supporting the notion that endocytosis is not required for HIV infection. In consequence of this apparent paradox, it has been suggested that both mechanisms, i.e. direct fusion to the plasma membrane and endocytosis, are functional pathways for HIV entry. I do not favor this position. Examination of the biology of CD4, its role in lymphocyte activation, and its function as the receptor for HIV imply a more active role for this molecule in virus infection than simply as a transient anchor for fusion events. The principal experiment of concern is the report by Maddon, et al., that human CD4- expressing murine cell lines remain resistant to infection by VSV pseudotypes bearing the HIV envelope glycoprotein. This paper and a subsequent review of HIV entry by Marsh and Dalgleish comment on the possibility that endocytosis of human CD4 may not occur in murine cells hence, the block to infectious entry of these pseudotype virions. Resolution of the mechanism for HIV infection is more than rationalization or semantics. The pathway of endocytosis presents an unique feature of cell biology. Should HIV infection be proven to occur via this mechanism, then appropriate assays for novel classes of antiviral agents could be developed. Other features of the endosomal vesicle, such as the unique composition of its membrane or the possible presence of specific proteolytic activities might play important roles in HIV infection and thus, present new targets for the development of antiviral agents. Indeed, the synthetic liposome AL721 is an effective inhibitor of HIV infection in vitro. This antiviral agent probably acts by decreasing the cholesterol content of the virion envelope. It was shown previously that the envelope is specifically enriched in cholesterol, a property not associated with the plasma membrane but clearly involved in Semliki Forest virus fusion within the endosome.(ABSTRACT TRUNCATED AT 400 WORDS)

Cell Line, Transformed↗
PubMed1991

Apoptosis as a mechanism of cell death in cultured T lymphoblasts acutely infected with HIV-1.

The mechanisms by which HIV-1 infection kills T lymphocytes are not clearly established. Apoptosis is an internally programmed cell death pathway that may regulate both T cell development and senescence, and that is characterized by cleavage of DNA at internucleosomal regions. The present experiments show that acute HIV-1 infection of MT2 lymphoblasts and activated normal peripheral blood mononuclear cells induces apoptosis. The addition of anti-gp120 neutralizing antibody, after HIV-1 infection of MT2 cells, permitted sustained high levels of viral replication, but blocked apoptosis and cell death. Apoptosis may account for the direct cytopathologic effects of HIV-1 in T cells.

CD4 Antigens↗
PubMed1990

Reinfection results in accumulation of unintegrated viral DNA in cytopathic and persistent human immunodeficiency virus type 1 infection of CEM cells.

High levels of unintegrated viral DNA accumulate during human immunodeficiency virus type 1 (HIV-1) infection of CEM T cells. Reinfection of already infected cells is required to attain these levels and reinfection also promotes the development of HIV-induced cytopathology. Rates of virus production, however, are independent of the accumulation of unintegrated viral DNA. Neutralizing antibody added soon after infection reduced viral DNA levels without appreciably affecting the production of cell-free viral p24 antigen or reverse transcriptase activity. Only 50 pM AZT were required to reduce the accumulation of unintegrated viral DNA by 50% in contrast to the 25 nM required to inhibit virus production by 50%. Cytopathology, as measured by number of syncytia in infected cell cultures, was correlated with highly elevated levels of unintegrated viral DNA. The minimal levels of unintegrated viral DNA present constitutively in the persistently infected HCEM cell line were consonant with the absence of cytopathic effects in these cells. These data demonstrate that inhibiting the reinfection of already infected cells modulates cytopathic HIV-1 infection to a form that is persistent and noncytopathic.

Antibodies, Monoclonal↗