Characterization of GP120 binding to CD4 and an assay that measures ability of sera to inhibit this binding

S. M. Schnittman, H. C. Lane, J. Roth, A. Burrows, T. M. Folks, J. H. Kehrl, S. Koenig, P. Berman, A. S. Fauci

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

There is evidence that the initial interaction between HIV-1 and the host that is essential for infection is the specific binding of the viral envelope glycoprotein, gp120, to the CD4 molecule found on certain T cells and monocytes. Most individuals infected with HIV develop antibodies against the gp120 protein. Although in vitro treatment of CD4+ T cells with mAb to a specific epitope of the CD4 molecule (T4a) blocks virus binding, syncytia formation, and infectivity, it is unclear if antibodies to gp120 from an infected individual that can inhibit the binding of gp120 to CD4 is in any way related to the clinical course of disease. Our present study characterizes the binding of 125I-labeled rgp120 to CD4+ cells, and describes an assay system that measures a potentially relevant form of immunity to HIV infection, i.e., the blocking of HIV binding to CD4+ cells. Optimal binding conditions included a 2-h incubation at 22°C, 4 x 106 CD4+ cells, and 1 nM gp120. The dissociation constant (K(D)) for gp120 binding to cell surface CD4 was 5 nM, and was inhibited by soluble CD4 and by mAb to T4a but not to T3 or T4. For the binding inhibition assay, negative controls included healthy seronegatives, seronegatives with connective tissue diseases, patients with HTLV-1 disease, and patients infected with HIV-2. In studying over 100 sera, the assay was highly sensitive (98%) and specific (100%). The majority of HIV+ sera could inhibit binding at dilutions of 1/100 to 1/1000. No correlation was noted between binding inhibition (BI) titer in this assay and clinical stage of HIV infection. In addition, there was no correlation between BI titer and HIV neutralizing activity. The BI titer was correlated with the titer of anti-gp160 (r = 0.63) and the titer of anti-gp120 (r = 0.52) antibodies determined by Western blot dilution. As with neutralizing antibodies and other forms of immune response to HIV, it is unclear what role antibody blocking of HIV binding to CD4+ cells may play in active immunity to HIV in infected individuals. This activity may prove to have some value in protection against initial HIV infection and, thus, the assay may be of use in monitoring vaccine trials.

Original languageEnglish (US)
Pages (from-to)4181-4186
Number of pages6
JournalJournal of Immunology
Volume141
Issue number12
StatePublished - 1988
Externally publishedYes

Fingerprint

HIV
HIV Infections
Serum
CD4 Antigens
HIV Antibodies
Active Immunity
T-Lymphocytes
Virus Attachment
HIV-2
Human T-lymphotropic virus 1
Blocking Antibodies
Connective Tissue Diseases
Antibodies
Giant Cells
Neutralizing Antibodies
HIV-1
Epitopes
Monocytes
Immunity
Glycoproteins

ASJC Scopus subject areas

  • Immunology

Cite this

Schnittman, S. M., Lane, H. C., Roth, J., Burrows, A., Folks, T. M., Kehrl, J. H., ... Fauci, A. S. (1988). Characterization of GP120 binding to CD4 and an assay that measures ability of sera to inhibit this binding. Journal of Immunology, 141(12), 4181-4186.

Characterization of GP120 binding to CD4 and an assay that measures ability of sera to inhibit this binding. / Schnittman, S. M.; Lane, H. C.; Roth, J.; Burrows, A.; Folks, T. M.; Kehrl, J. H.; Koenig, S.; Berman, P.; Fauci, A. S.

In: Journal of Immunology, Vol. 141, No. 12, 1988, p. 4181-4186.

Research output: Contribution to journalArticle

Schnittman, SM, Lane, HC, Roth, J, Burrows, A, Folks, TM, Kehrl, JH, Koenig, S, Berman, P & Fauci, AS 1988, 'Characterization of GP120 binding to CD4 and an assay that measures ability of sera to inhibit this binding', Journal of Immunology, vol. 141, no. 12, pp. 4181-4186.
Schnittman SM, Lane HC, Roth J, Burrows A, Folks TM, Kehrl JH et al. Characterization of GP120 binding to CD4 and an assay that measures ability of sera to inhibit this binding. Journal of Immunology. 1988;141(12):4181-4186.
Schnittman, S. M. ; Lane, H. C. ; Roth, J. ; Burrows, A. ; Folks, T. M. ; Kehrl, J. H. ; Koenig, S. ; Berman, P. ; Fauci, A. S. / Characterization of GP120 binding to CD4 and an assay that measures ability of sera to inhibit this binding. In: Journal of Immunology. 1988 ; Vol. 141, No. 12. pp. 4181-4186.
@article{9193de4d2604424b9ca17df57bc58af5,
title = "Characterization of GP120 binding to CD4 and an assay that measures ability of sera to inhibit this binding",
abstract = "There is evidence that the initial interaction between HIV-1 and the host that is essential for infection is the specific binding of the viral envelope glycoprotein, gp120, to the CD4 molecule found on certain T cells and monocytes. Most individuals infected with HIV develop antibodies against the gp120 protein. Although in vitro treatment of CD4+ T cells with mAb to a specific epitope of the CD4 molecule (T4a) blocks virus binding, syncytia formation, and infectivity, it is unclear if antibodies to gp120 from an infected individual that can inhibit the binding of gp120 to CD4 is in any way related to the clinical course of disease. Our present study characterizes the binding of 125I-labeled rgp120 to CD4+ cells, and describes an assay system that measures a potentially relevant form of immunity to HIV infection, i.e., the blocking of HIV binding to CD4+ cells. Optimal binding conditions included a 2-h incubation at 22°C, 4 x 106 CD4+ cells, and 1 nM gp120. The dissociation constant (K(D)) for gp120 binding to cell surface CD4 was 5 nM, and was inhibited by soluble CD4 and by mAb to T4a but not to T3 or T4. For the binding inhibition assay, negative controls included healthy seronegatives, seronegatives with connective tissue diseases, patients with HTLV-1 disease, and patients infected with HIV-2. In studying over 100 sera, the assay was highly sensitive (98{\%}) and specific (100{\%}). The majority of HIV+ sera could inhibit binding at dilutions of 1/100 to 1/1000. No correlation was noted between binding inhibition (BI) titer in this assay and clinical stage of HIV infection. In addition, there was no correlation between BI titer and HIV neutralizing activity. The BI titer was correlated with the titer of anti-gp160 (r = 0.63) and the titer of anti-gp120 (r = 0.52) antibodies determined by Western blot dilution. As with neutralizing antibodies and other forms of immune response to HIV, it is unclear what role antibody blocking of HIV binding to CD4+ cells may play in active immunity to HIV in infected individuals. This activity may prove to have some value in protection against initial HIV infection and, thus, the assay may be of use in monitoring vaccine trials.",
author = "Schnittman, {S. M.} and Lane, {H. C.} and J. Roth and A. Burrows and Folks, {T. M.} and Kehrl, {J. H.} and S. Koenig and P. Berman and Fauci, {A. S.}",
year = "1988",
language = "English (US)",
volume = "141",
pages = "4181--4186",
journal = "Journal of Immunology",
issn = "0022-1767",
publisher = "American Association of Immunologists",
number = "12",

}

TY - JOUR

T1 - Characterization of GP120 binding to CD4 and an assay that measures ability of sera to inhibit this binding

AU - Schnittman, S. M.

AU - Lane, H. C.

AU - Roth, J.

AU - Burrows, A.

AU - Folks, T. M.

AU - Kehrl, J. H.

AU - Koenig, S.

AU - Berman, P.

AU - Fauci, A. S.

PY - 1988

Y1 - 1988

N2 - There is evidence that the initial interaction between HIV-1 and the host that is essential for infection is the specific binding of the viral envelope glycoprotein, gp120, to the CD4 molecule found on certain T cells and monocytes. Most individuals infected with HIV develop antibodies against the gp120 protein. Although in vitro treatment of CD4+ T cells with mAb to a specific epitope of the CD4 molecule (T4a) blocks virus binding, syncytia formation, and infectivity, it is unclear if antibodies to gp120 from an infected individual that can inhibit the binding of gp120 to CD4 is in any way related to the clinical course of disease. Our present study characterizes the binding of 125I-labeled rgp120 to CD4+ cells, and describes an assay system that measures a potentially relevant form of immunity to HIV infection, i.e., the blocking of HIV binding to CD4+ cells. Optimal binding conditions included a 2-h incubation at 22°C, 4 x 106 CD4+ cells, and 1 nM gp120. The dissociation constant (K(D)) for gp120 binding to cell surface CD4 was 5 nM, and was inhibited by soluble CD4 and by mAb to T4a but not to T3 or T4. For the binding inhibition assay, negative controls included healthy seronegatives, seronegatives with connective tissue diseases, patients with HTLV-1 disease, and patients infected with HIV-2. In studying over 100 sera, the assay was highly sensitive (98%) and specific (100%). The majority of HIV+ sera could inhibit binding at dilutions of 1/100 to 1/1000. No correlation was noted between binding inhibition (BI) titer in this assay and clinical stage of HIV infection. In addition, there was no correlation between BI titer and HIV neutralizing activity. The BI titer was correlated with the titer of anti-gp160 (r = 0.63) and the titer of anti-gp120 (r = 0.52) antibodies determined by Western blot dilution. As with neutralizing antibodies and other forms of immune response to HIV, it is unclear what role antibody blocking of HIV binding to CD4+ cells may play in active immunity to HIV in infected individuals. This activity may prove to have some value in protection against initial HIV infection and, thus, the assay may be of use in monitoring vaccine trials.

AB - There is evidence that the initial interaction between HIV-1 and the host that is essential for infection is the specific binding of the viral envelope glycoprotein, gp120, to the CD4 molecule found on certain T cells and monocytes. Most individuals infected with HIV develop antibodies against the gp120 protein. Although in vitro treatment of CD4+ T cells with mAb to a specific epitope of the CD4 molecule (T4a) blocks virus binding, syncytia formation, and infectivity, it is unclear if antibodies to gp120 from an infected individual that can inhibit the binding of gp120 to CD4 is in any way related to the clinical course of disease. Our present study characterizes the binding of 125I-labeled rgp120 to CD4+ cells, and describes an assay system that measures a potentially relevant form of immunity to HIV infection, i.e., the blocking of HIV binding to CD4+ cells. Optimal binding conditions included a 2-h incubation at 22°C, 4 x 106 CD4+ cells, and 1 nM gp120. The dissociation constant (K(D)) for gp120 binding to cell surface CD4 was 5 nM, and was inhibited by soluble CD4 and by mAb to T4a but not to T3 or T4. For the binding inhibition assay, negative controls included healthy seronegatives, seronegatives with connective tissue diseases, patients with HTLV-1 disease, and patients infected with HIV-2. In studying over 100 sera, the assay was highly sensitive (98%) and specific (100%). The majority of HIV+ sera could inhibit binding at dilutions of 1/100 to 1/1000. No correlation was noted between binding inhibition (BI) titer in this assay and clinical stage of HIV infection. In addition, there was no correlation between BI titer and HIV neutralizing activity. The BI titer was correlated with the titer of anti-gp160 (r = 0.63) and the titer of anti-gp120 (r = 0.52) antibodies determined by Western blot dilution. As with neutralizing antibodies and other forms of immune response to HIV, it is unclear what role antibody blocking of HIV binding to CD4+ cells may play in active immunity to HIV in infected individuals. This activity may prove to have some value in protection against initial HIV infection and, thus, the assay may be of use in monitoring vaccine trials.

UR - http://www.scopus.com/inward/record.url?scp=0024207033&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024207033&partnerID=8YFLogxK

M3 - Article

C2 - 3264307

AN - SCOPUS:0024207033

VL - 141

SP - 4181

EP - 4186

JO - Journal of Immunology

JF - Journal of Immunology

SN - 0022-1767

IS - 12

ER -