Human immune response to HIV-1 Nef. II. Induction of HIV-1/HIV-2 Nef cross-reactive cytotoxic T lymphocytes in peripheral blood lymphocytes of non-infected healthy individuals

Abstract Several common metabolites and drugs in the serum in of patients with inflammatory, infectious, autoimmune, immunodeficient, neoplastic, and toxicant-induced diseases can produce artifactual suppression of the [methyl-3H]-thymidine assay, which is widely used to evaluate lymphocyte responsiveness. We have developed a sensitive, semiautomated, fluorescence-enhancement assay in which true immunosuppressors are measured in the presence of absence of such interfering substances. Peripheral blood lymphocytes are activated with mitogens in standard microtiter culture trays. Changes in lymphocyte DNA content are quantified with a reagent formulation containing mithramycin, the fluorescence of which is enhanced on binding to DNA in the presence of MgCl2. We solubilize cells within the intact microtiter tray by using an automated, inverted "Array Sonicator," and measure fluorescence with an automated, photon-counting fluorometer. With this system, immune response modulation can be accurately assessed in the presence of patients' sera and other complex test substances (e.g., supernates from hybridomas, fermentation vats, viral preparations, and macrophage cultures.

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High levels of HIV-1 replication show a clear correlation with downmodulation of Bcl-2 protein in peripheral blood lymphocytes of HIV-1-seropositive subjects

Journal of Medical Virology ◽

10.1002/(sici)1096-9071(199809)56:1<66::aid-jmv11>3.0.co;2-u ◽

1998 ◽

Vol 56 (1) ◽

pp. 66-73 ◽

Cited By ~ 10

Author(s):

Maria Carla Re ◽

Davide Gibellini ◽

Richard Aschbacher ◽

Monica Vignoli ◽

Giuliano Furlini ◽

...

Keyword(s):

Peripheral Blood ◽

Peripheral Blood Lymphocytes ◽

Clear Correlation ◽

Blood Lymphocytes ◽

Hiv 1

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Immune Response and Mitosis of Human Peripheral Blood Lymphocytes in vitro

Science ◽

10.1126/science.142.3596.1185 ◽

1963 ◽

Vol 142 (3596) ◽

pp. 1185-1187 ◽

Cited By ~ 247

Author(s):

K. Hirschhorn ◽

F. Bach ◽

R. L. Kolodny ◽

I. L. Firschein ◽

N. Hashem

Keyword(s):

Immune Response ◽

Peripheral Blood ◽

Human Peripheral Blood ◽

Peripheral Blood Lymphocytes ◽

Blood Lymphocytes ◽

Human Peripheral Blood Lymphocytes

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Expression and coreceptor activity of STRL33/Bonzo on primary peripheral blood lymphocytes

Blood ◽

10.1182/blood.v96.1.41 ◽

2000 ◽

Vol 96 (1) ◽

pp. 41-49 ◽

Cited By ~ 62

Author(s):

Matthew Sharron ◽

Stefan Pöhlmann ◽

Ken Price ◽

Elias Lolis ◽

Monica Tsang ◽

...

Keyword(s):

Rhesus Macaque ◽

Expression Pattern ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Interleukin 2 ◽

Peripheral Blood Lymphocytes ◽

Blood Lymphocytes ◽

Immunodeficiency Virus ◽

Hiv 1

Abstract CCR5 and CXCR4 are the major coreceptors that mediate human immunodeficiency virus 1 (HIV-1) infection, while most simian immunodeficiency virus (SIV) isolates use CCR5. A number of alternative coreceptors can also mediate infection of some virus strains in vitro, although little is known about their in vivo relevance. Therefore, we characterized the expression pattern and coreceptor activity of one of these alternative coreceptors, STRL33/Bonzo, using a newly developed monoclonal antibody. In addition to being highly expressed (approximately 1000-7000 STRL33 ABS [antibody binding sites]) on specific subsets of natural killer cells (CD3−/CD16−/low/CD56+ and CD3−/CD16low/CD56−) and CD19+ B lymphocytes (approximately 300-5000 STRL33 ABS), STRL33 was expressed at levels sufficient to support virus infection on freshly isolated, truly naive CD4+/CD45RA+/CD62L+cells (6000-11 000 ABS). STRL33 expression on peripheral blood mononuclear cells (PBMCs) was increased by mitogenic stimulation (OKT3/IL-2 [interleukin-2] had a greater effect than phytohemaglutinin (PHA)/IL-2), but it was dramatically decreased upon Ficoll purification. Infection of CCR5− human peripheral blood lymphocytes (PBLs) showed that 2 different SIV envelope (Env) proteins mediated entry into STRL33+cells. More importantly, the preferential infection of STRL33+ cells in CCR5− PBLs by an R5/X4/STRL33 HIV-1 maternal isolate in the presence of a potent CXCR4 antagonist (AMD3100) suggests that STRL33 can be used as a coreceptor by HIV-1 on primary cells. Rhesus macaque (rh) STRL33 was used less efficiently than human STRL33 by the majority of SIV Env proteins tested despite similar levels of expression, thereby making it less likely that STRL33 is a relevant coreceptor in the rhesus macaque system. In summary, the expression pattern and coreceptor activity of STRL33 suggest its involvement in trafficking of tumor-infiltrating lymphocytes and indicate that STRL33 may be a relevant coreceptor in vivo.

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Increased susceptibility to HIV-1 of peripheral blood lymphocytes in acute infection with Epstein-Barr virus

Journal of Medical Virology ◽

10.1002/jmv.10494 ◽

2003 ◽

Vol 71 (3) ◽

pp. 343-346 ◽

Cited By ~ 9

Author(s):

Masako Moriuchi ◽

Hiroyuki Moriuchi

Keyword(s):

Peripheral Blood ◽

Epstein Barr Virus ◽

Acute Infection ◽

Peripheral Blood Lymphocytes ◽

Blood Lymphocytes ◽

Barr Virus ◽

Epstein Barr ◽

Hiv 1 ◽

Increased Susceptibility

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Expression and coreceptor activity of STRL33/Bonzo on primary peripheral blood lymphocytes

Blood ◽

10.1182/blood.v96.1.41.013k53_41_49 ◽

2000 ◽

Vol 96 (1) ◽

pp. 41-49 ◽

Cited By ~ 4

Author(s):

Matthew Sharron ◽

Stefan Pöhlmann ◽

Ken Price ◽

Elias Lolis ◽

Monica Tsang ◽

...

Keyword(s):

Rhesus Macaque ◽

Expression Pattern ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Interleukin 2 ◽

Peripheral Blood Lymphocytes ◽

Blood Lymphocytes ◽

Immunodeficiency Virus ◽

Hiv 1

CCR5 and CXCR4 are the major coreceptors that mediate human immunodeficiency virus 1 (HIV-1) infection, while most simian immunodeficiency virus (SIV) isolates use CCR5. A number of alternative coreceptors can also mediate infection of some virus strains in vitro, although little is known about their in vivo relevance. Therefore, we characterized the expression pattern and coreceptor activity of one of these alternative coreceptors, STRL33/Bonzo, using a newly developed monoclonal antibody. In addition to being highly expressed (approximately 1000-7000 STRL33 ABS [antibody binding sites]) on specific subsets of natural killer cells (CD3−/CD16−/low/CD56+ and CD3−/CD16low/CD56−) and CD19+ B lymphocytes (approximately 300-5000 STRL33 ABS), STRL33 was expressed at levels sufficient to support virus infection on freshly isolated, truly naive CD4+/CD45RA+/CD62L+cells (6000-11 000 ABS). STRL33 expression on peripheral blood mononuclear cells (PBMCs) was increased by mitogenic stimulation (OKT3/IL-2 [interleukin-2] had a greater effect than phytohemaglutinin (PHA)/IL-2), but it was dramatically decreased upon Ficoll purification. Infection of CCR5− human peripheral blood lymphocytes (PBLs) showed that 2 different SIV envelope (Env) proteins mediated entry into STRL33+cells. More importantly, the preferential infection of STRL33+ cells in CCR5− PBLs by an R5/X4/STRL33 HIV-1 maternal isolate in the presence of a potent CXCR4 antagonist (AMD3100) suggests that STRL33 can be used as a coreceptor by HIV-1 on primary cells. Rhesus macaque (rh) STRL33 was used less efficiently than human STRL33 by the majority of SIV Env proteins tested despite similar levels of expression, thereby making it less likely that STRL33 is a relevant coreceptor in the rhesus macaque system. In summary, the expression pattern and coreceptor activity of STRL33 suggest its involvement in trafficking of tumor-infiltrating lymphocytes and indicate that STRL33 may be a relevant coreceptor in vivo.

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