Treatments for human immunodeficiency virus type 1 (HIV-1)-positive individuals that augment HIV-... more Treatments for human immunodeficiency virus type 1 (HIV-1)-positive individuals that augment HIV-1 suppression and have potential for achieving long-term control of HIV-1 viremia in the absence of antiretroviral therapy (ART) are urgently needed. We therefore conducted a phase I, clinical safety trial of a dendritic cell (DC)-based vaccination strategy as immunotherapy for HIV-1-positive individuals on ART. We studied 18 HIV-1-positive subjects on ART who underwent leukapheresis to obtain peripheral blood mononuclear cells for DC generation from monocytes cultured with cytokines. Mature DC were pulsed with three HIV-1 HLA*A0201 Gag, Env, and Pol peptides and one influenza A virus matrix protein peptide. The vaccine was administered to donors randomized to receive two vaccinations, either intravenously or subcutaneously. The primary end points were safety and tolerability of two doses of peptide-DC vaccine (3 million versus 10 million). Secondary end points included gamma interferon ...
Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc, 1994
A multiplex polymerase chain reaction (PCR)-liquid hybridization assay detecting a minimum 10 fg ... more A multiplex polymerase chain reaction (PCR)-liquid hybridization assay detecting a minimum 10 fg of cytomegaloviral (CMV) DNA has been developed. This assay successfully detected viral nucleic acids in 25/26 (96.2%) routinely embedded allograft liver biopsies with CMV hepatitis. No CMV DNA was detected in 15 native and 12 donor liver tissues sampled at the time of liver transplantation. These results indicate that archived formalin-fixed, paraffin-embedded liver tissues obtained from patients with significant liver disease can be successfully analyzed for CMV by PCR, retrospectively. Failure to detect CMV in native and donor livers implies that any latent virus present within these tissues is very focally distributed, or is below the threshold of detection applicable to PCR.
Cytomegalovirus (CMV) infection can cause severe disease and mortality in recipients of allogenei... more Cytomegalovirus (CMV) infection can cause severe disease and mortality in recipients of allogeneic bone marrow transplants (alloBMT) when either the donor or recipient is CMV seropositive (high-risk alloBMT). We investigated the efficacy of preemptive therapy guided by detection of CMV antigenemia. In 11 high-risk alloBMT recipients, high-dose ganciclovir (GCV) treatment was initiated at first positive antigenemia and was continued until antigenemia became negative. The treatment strategy prevented CMV disease during the follow-up period of the study in 7 alloBMT recipients with positive CMV antigenemia. Three other patients who were shown to be CMV antigenemia negative but positive for CMV DNA in blood by the polymerase chain reaction (PCR) were not treated and did not develop CMV disease. The eleventh patient was negative for CMV by all tests for the duration of the study and did not develop CMV disease. We have found antigenemia-guided preemptive GCV therapy to be an effective strategy for the prevention of CMV disease in high-risk alloBMT recipients.
Intestinal transplantation is a feasible therapy for patients with short-bowel syndrome. However,... more Intestinal transplantation is a feasible therapy for patients with short-bowel syndrome. However, cytomegalovirus (CMV) enteritis can cause complications. The aim of this study was to investigate the value of polymerase chain reaction (PCR)-based detection methods for CMV in the management of patients with small bowel transplants. Comparative evaluation of PCR with histopathology, shell-vial assay, and tube culture of intestinal biopsy specimens was used for the diagnosis of CMV enteritis in 21 patients. Ten patients experienced 21 episodes of CMV enteritis, diagnosed by histopathology, virology, or both. PCR had a sensitivity and specificity of 96% and 69%, respectively, compared with traditional methods, whereas the positive and negative predictive values were 35% and 99%, respectively. Three+ and 4+ signals corresponded to a specificity of 91% and positive predictive value of 59%, respectively. CMV was detected by PCR a median of 11 days (range, 0-32) earlier than other methods and lasted a median of 40 days (range, 21-80) in the 13 episodes, which became PCR-negative and in those patients who developed asymptomatic infection. In 8 episodes, CMV by PCR never became negative and was associated with a relapse of disease confirmed by other methods. PCR is a sensitive method for the early detection of CMV in intestinal biopsy specimens and can be used for preemptive therapy after intestinal transplantation.
The ability of dendritic cells (DC) to mediate CD4+ T cell help for cellular immunity is guided b... more The ability of dendritic cells (DC) to mediate CD4+ T cell help for cellular immunity is guided by instructive signals received during DC maturation, as well as the resulting pattern of DC responsiveness to the Th signal, CD40L. Furthermore, the professional transfer of antigenic information from migratory DC to lymph node–residing DC is critical for the effective induction of cellular immune responses. In this study we report that, in addition to their enhanced IL-12p70 producing capacity, human DC matured in the presence of inflammatory mediators of type 1 immunity are uniquely programmed to form networks of tunneling nanotube-like structures in response to CD40L-expressing Th cells or rCD40L. This immunologic process of DC reticulation facilitates intercellular trafficking of endosome-associated vesicles and Ag, but also pathogens such HIV-1, and is regulated by the opposing roles of IFN-γ and IL-4. The initiation of DC reticulation represents a novel helper function of CD40L and...
... The amount of HIV-1 DNA or RNA was calculated by comparing the signal intensity of the amplim... more ... The amount of HIV-1 DNA or RNA was calculated by comparing the signal intensity of the amplimer with a standard curve constructed by ... In this report, we describe a novel, internally controlled PCR (ICPCR) assay to quantitate HIV-1 Gag DNA and RNA in PBMC or plasma that ...
Clinical and diagnostic laboratory immunology, 1997
A reproducible, semiquantitative assay was developed to measure the level of the anti-human immun... more A reproducible, semiquantitative assay was developed to measure the level of the anti-human immunodeficiency virus type 1 (anti-HIV-1) suppressive activity of CD8+ T cells. The assay had a wide dynamic range and could be applied to a relatively small number of fresh and cryopreserved peripheral blood mononuclear cells and purified CD8+ T cells. The suppressive activity was not due to cytolytic activity and was not major histocompatibility complex class I restricted. Suppression of HIV-1 infection by CD8+ T cells was consistently demonstrable with both endogenously infected autologous CD4+ T cells and exogenously infected allogeneic CD4+ T cells. This assay can be used to monitor the level of antiviral activity of CD8+ T cells in a retrospective and prospective manner in studies of the natural history of HIV-1 infection and of subjects receiving anti-HIV-1 therapy and vaccines.
Journal of immunology (Baltimore, Md. : 1950), 1996
Dendritic cells are potent stimulators of Ag-specific T cell responses and have been implicated i... more Dendritic cells are potent stimulators of Ag-specific T cell responses and have been implicated in the pathogenesis of HIV-1 and other viral infections. Although cytokines may be involved in both of these processes, there is little information on the expression of cytokines by human blood dendritic cells. We characterized cytokine gene and protein expression in dendritic cells that were purified from normal human PBMC by flow cytometry and stimulated in vitro for up to 24 h with HIV-1 or herpes simplex virus (HSV). The unstimulated, uncultured dendritic cells were defined by their phenotype (HLA DR+ CD3- CD19- CD16- CD56- CD14-) and distinct morphology, lack of mRNA expression for CD3, CD14 and CD19, and presence of mRNA for CD4 and CD83. The purified dendritic cells also expressed CD4 (70-90%), CD33 (36-48%), and CD11c (44-54%); lacked CD1a expression (<1%); and were potent stimulators of an allogeneic MLR. The stimulated dendritic cells expressed mRNA for IFN-alpha, IL-1alpha, ...
Journal of immunology (Baltimore, Md. : 1950), 1996
The Ab-dependent cell-mediated cytotoxicity (ADCC) activity of anti-gp120 Abs in serum from four ... more The Ab-dependent cell-mediated cytotoxicity (ADCC) activity of anti-gp120 Abs in serum from four groups of HIV-1-positive individuals in the Multicenter AIDS Cohort Study was evaluated at several time points over a 10-yr period. HIV-1-positive individuals who progressed to AIDS within 3 yr of seroconversion (rapid progressors) were compared with seroconverters who did not progress to AIDS within 6 yr (nonrapid progressors) and individuals who were seropositive when they entered the study and did not progress to AIDS within 9-10 yr (nonprogressors). At the visit closest to AIDS, rapid progressors had significantly lower titers of Abs that mediate ADCC against HIV-1 gp120 than those of nonrapid progressors at corresponding visits or those of nonprogressors at any visit. Nonprogressors generally had high titers of ADCC Abs at all visits. Differences between ADCC titers of rapid progressors and nonrapid progressors or nonprogressors remained when longitudinal changes within individuals ...
Based on preclinical studies showing that CD8+ T lymphocytes of human immunodeficiency syndrome (... more Based on preclinical studies showing that CD8+ T lymphocytes of human immunodeficiency syndrome (HIV)-infected subjects have anti-HIV activities, a phase 1 study was undertaken to determine the safety and feasibility of infusing in vitro purified, activated, and expanded CD8+ cells as a therapeutic measure in seven patients with acquired immunodeficiency syndrome (AIDS)-related complex (ARC) or AIDS. Autologous CD8+ cells were first selectively isolated in monoclonal antibody-coated flasks from peripheral blood mononuclear cells recovered by leukapheresis. They were then cultured and expanded with phytohemagglutinin and recombinant interleukin-2 (rIL-2) before infusion. Five cycles of isolations and infusions of increasing numbers of CD8+ T cells were achieved in five of seven subjects. Five cycles could not be completed in two subjects with AIDS whose CD4+ cell counts were < or = 48/microliters. Infusions of CD8+ cells alone were well tolerated. Four patients received rIL-2 by c...
Cytolytic T lymphocytes play an important role in host defense against viral infections, includin... more Cytolytic T lymphocytes play an important role in host defense against viral infections, including human immunodeficiency virus (HIV). In a phase I clinical trial (protocol 080 of the AIDS Clinical Trials Group), generation of CD8+ effector cells from peripheral blood of patients with acquired immunodeficiency syndrome (AIDS)-related complex (ARC) or AIDS and safety of autologous adoptive transfer of these cells were evaluated. For therapeutic infusions, CD8+ T cells were purified by positive selection on anti-CD8 monoclonal antibody-coated flasks from leukapheresed peripheral blood of seven patients. These CD8+ T cells were cultured in the presence of interleukin-2 and phytohemagglutinin for up to 3 weeks to obtain cells sufficient for therapeutic infusions (10(8) to 10(10)). All 31 cell cultures established from the seven patients and used for therapy were highly enriched in CD8+ (mean, 97%), CD8+HLA-DR+ (50%), cytotoxic CD8+CD11b- (82%), and memory CD29+ (78%) T lymphocytes. In v...
The Centers for Disease Control reported in October 1991 that many people at risk for human immun... more The Centers for Disease Control reported in October 1991 that many people at risk for human immunodeficiency virus (HIV) infection had not been tested for antibodies to HIV. This study identifies differences among 110 gay and bisexual men in three small cities in Pennsylvania who decided whether to be tested for antibodies to HIV and, if so, whether to return for results. These men were given self-administered questionnaires and were offered free and confidential HIV antibody tests. Fifty percent of the men refused testing. Of those tested, only 35 percent returned to obtain test results. Contrary to other health prevention data, education was significantly and inversely related to being tested and to returning for results. Men who most often participated in the institutionalized gay community were least likely to be tested. The findings suggest that gay men who are most aware of the potential psychosocial problems associated with HIV antibody testing are more likely to avoid testing.
A total of 3,552 specimens for conventional cytomegalovirus (CMV) culture and shell vial assay fo... more A total of 3,552 specimens for conventional cytomegalovirus (CMV) culture and shell vial assay for CMV immediate-early antigen were obtained during a prospective randomized trial for prophylaxis of CMV disease after liver transplantation. Prophylaxis with ganciclovir for 2 weeks and then high-dose acyclovir for 2.5 months was compared with high-dose acyclovir alone for 3 months. During the first 12 weeks after transplantation, when the patients were on prophylaxis, there were significantly more clinical samples positive by the shell vial assay and negative by standard culture in comparison with the number of samples obtained from weeks 13 to 24, after prophylaxis was discontinued, that were positive by the shell vial assay and negative by standard culture. In contrast, significantly fewer samples were positive by both the shell vial assay and standard culture during the first 12 weeks compared with the number obtained 13 to 24 weeks after transplantation that were positive by both m...
Clinical and diagnostic laboratory immunology, 1995
Hepatitis C virus (HCV) infection may go undiagnosed and continue to present a source of communit... more Hepatitis C virus (HCV) infection may go undiagnosed and continue to present a source of community-acquired or transfusion-associated infection because of shortcomings in sensitivity, specificity, and reproducibility of serologic tests. This project was designed to longitudinally study persons who were HCV seropositive or were at risk for seroconversion to characterize the course of infection. Sequential serum samples obtained semiannually from 617 homosexual male volunteers were available for study from the Pittsburgh site of the Multicenter AIDS Cohort Study. Testing by anti-HCV enzyme immunoassay (EIA) was performed on baseline (1984 to 1985) and most-recent (censor date, August 1992) samples. Selected samples were also assayed for alanine aminotransferase and by recombinant immunoblot (RIBA II) and nested PCR. A total of 17 of 617 (2.8%) men were HCV seropositive at entry. Of the 600 seronegative men, 9 converted to HCV seropositive during the study interval. Parenteral sources ...
Clinical and diagnostic laboratory immunology, 1995
We previously reported that human immunodeficiency virus type 1 (HIV-1), herpes simplex virus (HS... more We previously reported that human immunodeficiency virus type 1 (HIV-1), herpes simplex virus (HSV), and Sendai virus induce higher levels of alpha interferon (IFN-alpha) in blood dendritic cells than in monocytes of healthy donors. In the present study, the levels of IFN-alpha induced by T-cell tropic (IIIb and RF) and monocytotropic (BaL) strains of HIV-1 and by HSV were significantly decreased in peripheral blood mononuclear cells (PBMCs) derived from subjects with asymptomatic and symptomatic HIV-1 infection. In contrast, Sendai virus, a paramyxovirus that induces proportionally more IFN-alpha in monocytes and B cells than do either HIV-1 or HSV, stimulated normal levels of IFN-alpha in PBMCs from the HIV-1-infected men. The IFN-alpha produced by PBMCs from the HIV-1-seropositive subjects was partially acid labile, whereas the IFN-alpha produced by PBMCs from the HIV-1-seronegative donors was acid stable. We hypothesize that there is a selective defect in IFN-alpha production by...
Lack of disease in long-term nonprogressors with human immunodeficiency virus type 1 (HIV-1) infe... more Lack of disease in long-term nonprogressors with human immunodeficiency virus type 1 (HIV-1) infection was strongly associated with very low copy numbers of HIV-1 DNA and RNA in peripheral blood mononuclear cells and plasma and the presence of high levels of anti-HIV-1 CD8+ memory cytotoxic T lymphocytes specific for Gag, Pol, and Env, compared with levels present in intermediate and advanced progressors. CD8+ memory cytotoxic T lymphocytes may have an important role in controlling HIV-1 replication and preventing disease in long-term nonprogressors.
Treatments for human immunodeficiency virus type 1 (HIV-1)-positive individuals that augment HIV-... more Treatments for human immunodeficiency virus type 1 (HIV-1)-positive individuals that augment HIV-1 suppression and have potential for achieving long-term control of HIV-1 viremia in the absence of antiretroviral therapy (ART) are urgently needed. We therefore conducted a phase I, clinical safety trial of a dendritic cell (DC)-based vaccination strategy as immunotherapy for HIV-1-positive individuals on ART. We studied 18 HIV-1-positive subjects on ART who underwent leukapheresis to obtain peripheral blood mononuclear cells for DC generation from monocytes cultured with cytokines. Mature DC were pulsed with three HIV-1 HLA*A0201 Gag, Env, and Pol peptides and one influenza A virus matrix protein peptide. The vaccine was administered to donors randomized to receive two vaccinations, either intravenously or subcutaneously. The primary end points were safety and tolerability of two doses of peptide-DC vaccine (3 million versus 10 million). Secondary end points included gamma interferon ...
Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc, 1994
A multiplex polymerase chain reaction (PCR)-liquid hybridization assay detecting a minimum 10 fg ... more A multiplex polymerase chain reaction (PCR)-liquid hybridization assay detecting a minimum 10 fg of cytomegaloviral (CMV) DNA has been developed. This assay successfully detected viral nucleic acids in 25/26 (96.2%) routinely embedded allograft liver biopsies with CMV hepatitis. No CMV DNA was detected in 15 native and 12 donor liver tissues sampled at the time of liver transplantation. These results indicate that archived formalin-fixed, paraffin-embedded liver tissues obtained from patients with significant liver disease can be successfully analyzed for CMV by PCR, retrospectively. Failure to detect CMV in native and donor livers implies that any latent virus present within these tissues is very focally distributed, or is below the threshold of detection applicable to PCR.
Cytomegalovirus (CMV) infection can cause severe disease and mortality in recipients of allogenei... more Cytomegalovirus (CMV) infection can cause severe disease and mortality in recipients of allogeneic bone marrow transplants (alloBMT) when either the donor or recipient is CMV seropositive (high-risk alloBMT). We investigated the efficacy of preemptive therapy guided by detection of CMV antigenemia. In 11 high-risk alloBMT recipients, high-dose ganciclovir (GCV) treatment was initiated at first positive antigenemia and was continued until antigenemia became negative. The treatment strategy prevented CMV disease during the follow-up period of the study in 7 alloBMT recipients with positive CMV antigenemia. Three other patients who were shown to be CMV antigenemia negative but positive for CMV DNA in blood by the polymerase chain reaction (PCR) were not treated and did not develop CMV disease. The eleventh patient was negative for CMV by all tests for the duration of the study and did not develop CMV disease. We have found antigenemia-guided preemptive GCV therapy to be an effective strategy for the prevention of CMV disease in high-risk alloBMT recipients.
Intestinal transplantation is a feasible therapy for patients with short-bowel syndrome. However,... more Intestinal transplantation is a feasible therapy for patients with short-bowel syndrome. However, cytomegalovirus (CMV) enteritis can cause complications. The aim of this study was to investigate the value of polymerase chain reaction (PCR)-based detection methods for CMV in the management of patients with small bowel transplants. Comparative evaluation of PCR with histopathology, shell-vial assay, and tube culture of intestinal biopsy specimens was used for the diagnosis of CMV enteritis in 21 patients. Ten patients experienced 21 episodes of CMV enteritis, diagnosed by histopathology, virology, or both. PCR had a sensitivity and specificity of 96% and 69%, respectively, compared with traditional methods, whereas the positive and negative predictive values were 35% and 99%, respectively. Three+ and 4+ signals corresponded to a specificity of 91% and positive predictive value of 59%, respectively. CMV was detected by PCR a median of 11 days (range, 0-32) earlier than other methods and lasted a median of 40 days (range, 21-80) in the 13 episodes, which became PCR-negative and in those patients who developed asymptomatic infection. In 8 episodes, CMV by PCR never became negative and was associated with a relapse of disease confirmed by other methods. PCR is a sensitive method for the early detection of CMV in intestinal biopsy specimens and can be used for preemptive therapy after intestinal transplantation.
The ability of dendritic cells (DC) to mediate CD4+ T cell help for cellular immunity is guided b... more The ability of dendritic cells (DC) to mediate CD4+ T cell help for cellular immunity is guided by instructive signals received during DC maturation, as well as the resulting pattern of DC responsiveness to the Th signal, CD40L. Furthermore, the professional transfer of antigenic information from migratory DC to lymph node–residing DC is critical for the effective induction of cellular immune responses. In this study we report that, in addition to their enhanced IL-12p70 producing capacity, human DC matured in the presence of inflammatory mediators of type 1 immunity are uniquely programmed to form networks of tunneling nanotube-like structures in response to CD40L-expressing Th cells or rCD40L. This immunologic process of DC reticulation facilitates intercellular trafficking of endosome-associated vesicles and Ag, but also pathogens such HIV-1, and is regulated by the opposing roles of IFN-γ and IL-4. The initiation of DC reticulation represents a novel helper function of CD40L and...
... The amount of HIV-1 DNA or RNA was calculated by comparing the signal intensity of the amplim... more ... The amount of HIV-1 DNA or RNA was calculated by comparing the signal intensity of the amplimer with a standard curve constructed by ... In this report, we describe a novel, internally controlled PCR (ICPCR) assay to quantitate HIV-1 Gag DNA and RNA in PBMC or plasma that ...
Clinical and diagnostic laboratory immunology, 1997
A reproducible, semiquantitative assay was developed to measure the level of the anti-human immun... more A reproducible, semiquantitative assay was developed to measure the level of the anti-human immunodeficiency virus type 1 (anti-HIV-1) suppressive activity of CD8+ T cells. The assay had a wide dynamic range and could be applied to a relatively small number of fresh and cryopreserved peripheral blood mononuclear cells and purified CD8+ T cells. The suppressive activity was not due to cytolytic activity and was not major histocompatibility complex class I restricted. Suppression of HIV-1 infection by CD8+ T cells was consistently demonstrable with both endogenously infected autologous CD4+ T cells and exogenously infected allogeneic CD4+ T cells. This assay can be used to monitor the level of antiviral activity of CD8+ T cells in a retrospective and prospective manner in studies of the natural history of HIV-1 infection and of subjects receiving anti-HIV-1 therapy and vaccines.
Journal of immunology (Baltimore, Md. : 1950), 1996
Dendritic cells are potent stimulators of Ag-specific T cell responses and have been implicated i... more Dendritic cells are potent stimulators of Ag-specific T cell responses and have been implicated in the pathogenesis of HIV-1 and other viral infections. Although cytokines may be involved in both of these processes, there is little information on the expression of cytokines by human blood dendritic cells. We characterized cytokine gene and protein expression in dendritic cells that were purified from normal human PBMC by flow cytometry and stimulated in vitro for up to 24 h with HIV-1 or herpes simplex virus (HSV). The unstimulated, uncultured dendritic cells were defined by their phenotype (HLA DR+ CD3- CD19- CD16- CD56- CD14-) and distinct morphology, lack of mRNA expression for CD3, CD14 and CD19, and presence of mRNA for CD4 and CD83. The purified dendritic cells also expressed CD4 (70-90%), CD33 (36-48%), and CD11c (44-54%); lacked CD1a expression (<1%); and were potent stimulators of an allogeneic MLR. The stimulated dendritic cells expressed mRNA for IFN-alpha, IL-1alpha, ...
Journal of immunology (Baltimore, Md. : 1950), 1996
The Ab-dependent cell-mediated cytotoxicity (ADCC) activity of anti-gp120 Abs in serum from four ... more The Ab-dependent cell-mediated cytotoxicity (ADCC) activity of anti-gp120 Abs in serum from four groups of HIV-1-positive individuals in the Multicenter AIDS Cohort Study was evaluated at several time points over a 10-yr period. HIV-1-positive individuals who progressed to AIDS within 3 yr of seroconversion (rapid progressors) were compared with seroconverters who did not progress to AIDS within 6 yr (nonrapid progressors) and individuals who were seropositive when they entered the study and did not progress to AIDS within 9-10 yr (nonprogressors). At the visit closest to AIDS, rapid progressors had significantly lower titers of Abs that mediate ADCC against HIV-1 gp120 than those of nonrapid progressors at corresponding visits or those of nonprogressors at any visit. Nonprogressors generally had high titers of ADCC Abs at all visits. Differences between ADCC titers of rapid progressors and nonrapid progressors or nonprogressors remained when longitudinal changes within individuals ...
Based on preclinical studies showing that CD8+ T lymphocytes of human immunodeficiency syndrome (... more Based on preclinical studies showing that CD8+ T lymphocytes of human immunodeficiency syndrome (HIV)-infected subjects have anti-HIV activities, a phase 1 study was undertaken to determine the safety and feasibility of infusing in vitro purified, activated, and expanded CD8+ cells as a therapeutic measure in seven patients with acquired immunodeficiency syndrome (AIDS)-related complex (ARC) or AIDS. Autologous CD8+ cells were first selectively isolated in monoclonal antibody-coated flasks from peripheral blood mononuclear cells recovered by leukapheresis. They were then cultured and expanded with phytohemagglutinin and recombinant interleukin-2 (rIL-2) before infusion. Five cycles of isolations and infusions of increasing numbers of CD8+ T cells were achieved in five of seven subjects. Five cycles could not be completed in two subjects with AIDS whose CD4+ cell counts were < or = 48/microliters. Infusions of CD8+ cells alone were well tolerated. Four patients received rIL-2 by c...
Cytolytic T lymphocytes play an important role in host defense against viral infections, includin... more Cytolytic T lymphocytes play an important role in host defense against viral infections, including human immunodeficiency virus (HIV). In a phase I clinical trial (protocol 080 of the AIDS Clinical Trials Group), generation of CD8+ effector cells from peripheral blood of patients with acquired immunodeficiency syndrome (AIDS)-related complex (ARC) or AIDS and safety of autologous adoptive transfer of these cells were evaluated. For therapeutic infusions, CD8+ T cells were purified by positive selection on anti-CD8 monoclonal antibody-coated flasks from leukapheresed peripheral blood of seven patients. These CD8+ T cells were cultured in the presence of interleukin-2 and phytohemagglutinin for up to 3 weeks to obtain cells sufficient for therapeutic infusions (10(8) to 10(10)). All 31 cell cultures established from the seven patients and used for therapy were highly enriched in CD8+ (mean, 97%), CD8+HLA-DR+ (50%), cytotoxic CD8+CD11b- (82%), and memory CD29+ (78%) T lymphocytes. In v...
The Centers for Disease Control reported in October 1991 that many people at risk for human immun... more The Centers for Disease Control reported in October 1991 that many people at risk for human immunodeficiency virus (HIV) infection had not been tested for antibodies to HIV. This study identifies differences among 110 gay and bisexual men in three small cities in Pennsylvania who decided whether to be tested for antibodies to HIV and, if so, whether to return for results. These men were given self-administered questionnaires and were offered free and confidential HIV antibody tests. Fifty percent of the men refused testing. Of those tested, only 35 percent returned to obtain test results. Contrary to other health prevention data, education was significantly and inversely related to being tested and to returning for results. Men who most often participated in the institutionalized gay community were least likely to be tested. The findings suggest that gay men who are most aware of the potential psychosocial problems associated with HIV antibody testing are more likely to avoid testing.
A total of 3,552 specimens for conventional cytomegalovirus (CMV) culture and shell vial assay fo... more A total of 3,552 specimens for conventional cytomegalovirus (CMV) culture and shell vial assay for CMV immediate-early antigen were obtained during a prospective randomized trial for prophylaxis of CMV disease after liver transplantation. Prophylaxis with ganciclovir for 2 weeks and then high-dose acyclovir for 2.5 months was compared with high-dose acyclovir alone for 3 months. During the first 12 weeks after transplantation, when the patients were on prophylaxis, there were significantly more clinical samples positive by the shell vial assay and negative by standard culture in comparison with the number of samples obtained from weeks 13 to 24, after prophylaxis was discontinued, that were positive by the shell vial assay and negative by standard culture. In contrast, significantly fewer samples were positive by both the shell vial assay and standard culture during the first 12 weeks compared with the number obtained 13 to 24 weeks after transplantation that were positive by both m...
Clinical and diagnostic laboratory immunology, 1995
Hepatitis C virus (HCV) infection may go undiagnosed and continue to present a source of communit... more Hepatitis C virus (HCV) infection may go undiagnosed and continue to present a source of community-acquired or transfusion-associated infection because of shortcomings in sensitivity, specificity, and reproducibility of serologic tests. This project was designed to longitudinally study persons who were HCV seropositive or were at risk for seroconversion to characterize the course of infection. Sequential serum samples obtained semiannually from 617 homosexual male volunteers were available for study from the Pittsburgh site of the Multicenter AIDS Cohort Study. Testing by anti-HCV enzyme immunoassay (EIA) was performed on baseline (1984 to 1985) and most-recent (censor date, August 1992) samples. Selected samples were also assayed for alanine aminotransferase and by recombinant immunoblot (RIBA II) and nested PCR. A total of 17 of 617 (2.8%) men were HCV seropositive at entry. Of the 600 seronegative men, 9 converted to HCV seropositive during the study interval. Parenteral sources ...
Clinical and diagnostic laboratory immunology, 1995
We previously reported that human immunodeficiency virus type 1 (HIV-1), herpes simplex virus (HS... more We previously reported that human immunodeficiency virus type 1 (HIV-1), herpes simplex virus (HSV), and Sendai virus induce higher levels of alpha interferon (IFN-alpha) in blood dendritic cells than in monocytes of healthy donors. In the present study, the levels of IFN-alpha induced by T-cell tropic (IIIb and RF) and monocytotropic (BaL) strains of HIV-1 and by HSV were significantly decreased in peripheral blood mononuclear cells (PBMCs) derived from subjects with asymptomatic and symptomatic HIV-1 infection. In contrast, Sendai virus, a paramyxovirus that induces proportionally more IFN-alpha in monocytes and B cells than do either HIV-1 or HSV, stimulated normal levels of IFN-alpha in PBMCs from the HIV-1-infected men. The IFN-alpha produced by PBMCs from the HIV-1-seropositive subjects was partially acid labile, whereas the IFN-alpha produced by PBMCs from the HIV-1-seronegative donors was acid stable. We hypothesize that there is a selective defect in IFN-alpha production by...
Lack of disease in long-term nonprogressors with human immunodeficiency virus type 1 (HIV-1) infe... more Lack of disease in long-term nonprogressors with human immunodeficiency virus type 1 (HIV-1) infection was strongly associated with very low copy numbers of HIV-1 DNA and RNA in peripheral blood mononuclear cells and plasma and the presence of high levels of anti-HIV-1 CD8+ memory cytotoxic T lymphocytes specific for Gag, Pol, and Env, compared with levels present in intermediate and advanced progressors. CD8+ memory cytotoxic T lymphocytes may have an important role in controlling HIV-1 replication and preventing disease in long-term nonprogressors.
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