Development of a monoclonal antibody-based p24 capsid antigen detection assay for HTLV-I, HTLV-II, and STLV-I infection

G. Toedter, S. Pearlman, D. Hofheinz, J. Blakeslee, G. Cockerell, C. Dezzutti, J. Yee, R. B. Lal, Michael Dale Lairmore

Research output: Contribution to journalArticle

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Abstract

A monoclonal antibody-based antigen capture enzyme-linked immunosorbent assay (ELISA) was developed and employed to detect p24 capsid antigen from human T-cell lymphotropic viruses type I and II (HTLV-I, HTLV-II), simian T- cell lymphotropic virus type I (STLV-I) -infected cell lines, and from mononuclear cell cocultures of HTLV-infected humans and STLV-I infected monkeys. A monoclonal antibody specific for HTLV p24 and p53 capsid antigens was coated onto 96-well microtiter plates to capture HTLV/STLV antigen. Captured antigen was then detected by the addition of a polyclonal, biotinylated human anti-HTLV-I antibody, and color developed with tetramethyl benzidine/H2O2 substrate. As little as 15 pg/ml of HTLV-I p24 antigen could be detected in this assay. Culture supernatants from HTLV-I-infected cell lines (HUT-102, MT-2, C5/MJ), HTLV-II-infected cell lines (Mo-T, Mo-B, PanG 12.1, NRA) and STLV-I-infected cell lines (Matsu, NEPC M39) were all positive in the assay. In addition, p24 was detected from peripheral blood mononuclear cell (PBMC) cocultures of 8 (100%) HTLV-I diseased patients, 14 of 20 (70%) HTLV-I and HTLV-II-infected, asymptomatic persons, and 8 of 8 (100%) STLV-I- infected, asymptomatic monkeys. Culture supernatants of cells infected with human immunodeficiency virus type (HIV-1), simian immunodeficiency virus (SIV), Chlamydia trachomatis, cytomegalovirus (CMV), herpes simplex I and II (HSV), feline leukemia virus (FELV), bovine leukemia virus (BLV), and bovine immunodeficiency virus (BIV) were all negative. Similarly, normal human peripheral blood mononuclear cells and uninfected, transformed human T cells, were also negative in the assay. The antigen assay is a sensitive and specific method for the detection of HTLV-I, HTLV-II, and STLV-I p24 capsid antigen and can replace reverse transcriptase assays in the confirmation of tissue cultures for these retroviruses.

Original languageEnglish (US)
Pages (from-to)527-532
Number of pages6
JournalAIDS Research and Human Retroviruses
Volume8
Issue number4
StatePublished - 1992
Externally publishedYes

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Simian T-lymphotropic virus 1
Human T-lymphotropic virus 2
Human T-lymphotropic virus 1
Capsid
Virus Diseases
Monoclonal Antibodies
T-Lymphocytes
Antigens
Cell Line
Coculture Techniques
Deltaretrovirus Antigens
HTLV-I Antigens
Haplorhini
HIV-1
Blood Cells
Bovine Immunodeficiency Virus
HTLV-I Antibodies
Bovine Leukemia Virus
Feline Leukemia Virus
Tumor Suppressor Protein p53

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

Development of a monoclonal antibody-based p24 capsid antigen detection assay for HTLV-I, HTLV-II, and STLV-I infection. / Toedter, G.; Pearlman, S.; Hofheinz, D.; Blakeslee, J.; Cockerell, G.; Dezzutti, C.; Yee, J.; Lal, R. B.; Lairmore, Michael Dale.

In: AIDS Research and Human Retroviruses, Vol. 8, No. 4, 1992, p. 527-532.

Research output: Contribution to journalArticle

Toedter, G, Pearlman, S, Hofheinz, D, Blakeslee, J, Cockerell, G, Dezzutti, C, Yee, J, Lal, RB & Lairmore, MD 1992, 'Development of a monoclonal antibody-based p24 capsid antigen detection assay for HTLV-I, HTLV-II, and STLV-I infection', AIDS Research and Human Retroviruses, vol. 8, no. 4, pp. 527-532.
Toedter, G. ; Pearlman, S. ; Hofheinz, D. ; Blakeslee, J. ; Cockerell, G. ; Dezzutti, C. ; Yee, J. ; Lal, R. B. ; Lairmore, Michael Dale. / Development of a monoclonal antibody-based p24 capsid antigen detection assay for HTLV-I, HTLV-II, and STLV-I infection. In: AIDS Research and Human Retroviruses. 1992 ; Vol. 8, No. 4. pp. 527-532.
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abstract = "A monoclonal antibody-based antigen capture enzyme-linked immunosorbent assay (ELISA) was developed and employed to detect p24 capsid antigen from human T-cell lymphotropic viruses type I and II (HTLV-I, HTLV-II), simian T- cell lymphotropic virus type I (STLV-I) -infected cell lines, and from mononuclear cell cocultures of HTLV-infected humans and STLV-I infected monkeys. A monoclonal antibody specific for HTLV p24 and p53 capsid antigens was coated onto 96-well microtiter plates to capture HTLV/STLV antigen. Captured antigen was then detected by the addition of a polyclonal, biotinylated human anti-HTLV-I antibody, and color developed with tetramethyl benzidine/H2O2 substrate. As little as 15 pg/ml of HTLV-I p24 antigen could be detected in this assay. Culture supernatants from HTLV-I-infected cell lines (HUT-102, MT-2, C5/MJ), HTLV-II-infected cell lines (Mo-T, Mo-B, PanG 12.1, NRA) and STLV-I-infected cell lines (Matsu, NEPC M39) were all positive in the assay. In addition, p24 was detected from peripheral blood mononuclear cell (PBMC) cocultures of 8 (100{\%}) HTLV-I diseased patients, 14 of 20 (70{\%}) HTLV-I and HTLV-II-infected, asymptomatic persons, and 8 of 8 (100{\%}) STLV-I- infected, asymptomatic monkeys. Culture supernatants of cells infected with human immunodeficiency virus type (HIV-1), simian immunodeficiency virus (SIV), Chlamydia trachomatis, cytomegalovirus (CMV), herpes simplex I and II (HSV), feline leukemia virus (FELV), bovine leukemia virus (BLV), and bovine immunodeficiency virus (BIV) were all negative. Similarly, normal human peripheral blood mononuclear cells and uninfected, transformed human T cells, were also negative in the assay. The antigen assay is a sensitive and specific method for the detection of HTLV-I, HTLV-II, and STLV-I p24 capsid antigen and can replace reverse transcriptase assays in the confirmation of tissue cultures for these retroviruses.",
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T1 - Development of a monoclonal antibody-based p24 capsid antigen detection assay for HTLV-I, HTLV-II, and STLV-I infection

AU - Toedter, G.

AU - Pearlman, S.

AU - Hofheinz, D.

AU - Blakeslee, J.

AU - Cockerell, G.

AU - Dezzutti, C.

AU - Yee, J.

AU - Lal, R. B.

AU - Lairmore, Michael Dale

PY - 1992

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N2 - A monoclonal antibody-based antigen capture enzyme-linked immunosorbent assay (ELISA) was developed and employed to detect p24 capsid antigen from human T-cell lymphotropic viruses type I and II (HTLV-I, HTLV-II), simian T- cell lymphotropic virus type I (STLV-I) -infected cell lines, and from mononuclear cell cocultures of HTLV-infected humans and STLV-I infected monkeys. A monoclonal antibody specific for HTLV p24 and p53 capsid antigens was coated onto 96-well microtiter plates to capture HTLV/STLV antigen. Captured antigen was then detected by the addition of a polyclonal, biotinylated human anti-HTLV-I antibody, and color developed with tetramethyl benzidine/H2O2 substrate. As little as 15 pg/ml of HTLV-I p24 antigen could be detected in this assay. Culture supernatants from HTLV-I-infected cell lines (HUT-102, MT-2, C5/MJ), HTLV-II-infected cell lines (Mo-T, Mo-B, PanG 12.1, NRA) and STLV-I-infected cell lines (Matsu, NEPC M39) were all positive in the assay. In addition, p24 was detected from peripheral blood mononuclear cell (PBMC) cocultures of 8 (100%) HTLV-I diseased patients, 14 of 20 (70%) HTLV-I and HTLV-II-infected, asymptomatic persons, and 8 of 8 (100%) STLV-I- infected, asymptomatic monkeys. Culture supernatants of cells infected with human immunodeficiency virus type (HIV-1), simian immunodeficiency virus (SIV), Chlamydia trachomatis, cytomegalovirus (CMV), herpes simplex I and II (HSV), feline leukemia virus (FELV), bovine leukemia virus (BLV), and bovine immunodeficiency virus (BIV) were all negative. Similarly, normal human peripheral blood mononuclear cells and uninfected, transformed human T cells, were also negative in the assay. The antigen assay is a sensitive and specific method for the detection of HTLV-I, HTLV-II, and STLV-I p24 capsid antigen and can replace reverse transcriptase assays in the confirmation of tissue cultures for these retroviruses.

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