Detection and isolation of type C retrovirus particles from fresh and cultured lymphocytes of a patient with cutaneous T-cell lymphoma

B. J. Poiesz, F. W. Ruscetti, A. F. Gazdar, P. A. Bunn, J. D. Minna, R. C. Gallo

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Abstract

Retrovirus particles with type C morphology were found in two T-cell lymphoblastoid cell lines, HUT 102 and CTCL-3, and in fresh peripheral blood lymphocytes obtained from a patient with a cutaneous T-cell lymphoma (mycosis fungoides). The cell lines continuously produce these viruses, which are collectively referred to as HTLV, strain CR (HTLV[CR]). Originally, the production of virus from HUT 102 cells required induction with 5-iodo-2'-deoxyuridine, but the cell line became a constitutive producer of virus at its 56th passage. Cell line CTCL-3 has been a constitutive producer of virus from its second passage in culture. Both mature and immature extracellular virus particles were seen in thin-section electron micrographs of fixed, pelleted cellular material; on occasion, typical type C budding virus particles were seen. No form of intracellular virus particle has been seen. Mature particles were 100-110 nm in diameter, consisted of an electron-dense core surrounded by an outer membrane separated by an electron-lucent region, banded at a density of 1.16 g/ml on a continuous 25-65% sucrose gradient, and contained 70S RNA and a DNA polymerase activity typical of viral reverse transcriptase (RT;RNA-dependent DNA nucleotidyltransferase). Under certain conditions of assay, HTLV[CR] RT showed cation preference for Mg 2+ over Mn 2+, distinct from the characteristics of cellular DNA polymerases purified from human lymphocytes and the RT from most type C viruses. Antibodies to cellular DNA polymerase γ and antibodies against RT purified from several animal retroviruses failed to detectably interact with HTLV[CR]RT under conditions that were positive for the respective homologous DNA polymerase, demonstrating a lack of close relationship of HTLV[CR]RT to cellular DNA polymerases γ or RT of these viruses. Six major proteins, with sizes of approximately 10,000, 13,000, 19,000, 24,000, 42,000 and 52,000 daltons were apparent when doubly banded, disrupted HTLV[CR] particles were chromatographed on a NaDodSO 4/polyacrylamide gel. The number of these particle-associated proteins is consistent with the expected proteins of a retrovirus, but the sizes of some are distinct from those of most known retroviruses of the primate subgroups.

Original languageEnglish (US)
Pages (from-to)7415-7419
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume77
Issue number12 II
DOIs
StatePublished - 1980

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Cutaneous T-Cell Lymphoma
Retroviridae
DNA-Directed DNA Polymerase
Lymphocytes
Viruses
Virion
Cell Line
Electrons
DNA Nucleotidyltransferases
Retroviridae Proteins
RNA
Idoxuridine
Mycosis Fungoides
Antibodies
RNA-Directed DNA Polymerase
Primates
Sucrose
Cations
Proteins
T-Lymphocytes

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

@article{a3269547e8784597ad81c1d880790870,
title = "Detection and isolation of type C retrovirus particles from fresh and cultured lymphocytes of a patient with cutaneous T-cell lymphoma",
abstract = "Retrovirus particles with type C morphology were found in two T-cell lymphoblastoid cell lines, HUT 102 and CTCL-3, and in fresh peripheral blood lymphocytes obtained from a patient with a cutaneous T-cell lymphoma (mycosis fungoides). The cell lines continuously produce these viruses, which are collectively referred to as HTLV, strain CR (HTLV[CR]). Originally, the production of virus from HUT 102 cells required induction with 5-iodo-2'-deoxyuridine, but the cell line became a constitutive producer of virus at its 56th passage. Cell line CTCL-3 has been a constitutive producer of virus from its second passage in culture. Both mature and immature extracellular virus particles were seen in thin-section electron micrographs of fixed, pelleted cellular material; on occasion, typical type C budding virus particles were seen. No form of intracellular virus particle has been seen. Mature particles were 100-110 nm in diameter, consisted of an electron-dense core surrounded by an outer membrane separated by an electron-lucent region, banded at a density of 1.16 g/ml on a continuous 25-65{\%} sucrose gradient, and contained 70S RNA and a DNA polymerase activity typical of viral reverse transcriptase (RT;RNA-dependent DNA nucleotidyltransferase). Under certain conditions of assay, HTLV[CR] RT showed cation preference for Mg 2+ over Mn 2+, distinct from the characteristics of cellular DNA polymerases purified from human lymphocytes and the RT from most type C viruses. Antibodies to cellular DNA polymerase γ and antibodies against RT purified from several animal retroviruses failed to detectably interact with HTLV[CR]RT under conditions that were positive for the respective homologous DNA polymerase, demonstrating a lack of close relationship of HTLV[CR]RT to cellular DNA polymerases γ or RT of these viruses. Six major proteins, with sizes of approximately 10,000, 13,000, 19,000, 24,000, 42,000 and 52,000 daltons were apparent when doubly banded, disrupted HTLV[CR] particles were chromatographed on a NaDodSO 4/polyacrylamide gel. The number of these particle-associated proteins is consistent with the expected proteins of a retrovirus, but the sizes of some are distinct from those of most known retroviruses of the primate subgroups.",
author = "Poiesz, {B. J.} and Ruscetti, {F. W.} and Gazdar, {A. F.} and Bunn, {P. A.} and Minna, {J. D.} and Gallo, {R. C.}",
year = "1980",
doi = "10.1073/pnas.77.12.7415",
language = "English (US)",
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journal = "Proceedings of the National Academy of Sciences of the United States of America",
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T1 - Detection and isolation of type C retrovirus particles from fresh and cultured lymphocytes of a patient with cutaneous T-cell lymphoma

AU - Poiesz, B. J.

AU - Ruscetti, F. W.

AU - Gazdar, A. F.

AU - Bunn, P. A.

AU - Minna, J. D.

AU - Gallo, R. C.

PY - 1980

Y1 - 1980

N2 - Retrovirus particles with type C morphology were found in two T-cell lymphoblastoid cell lines, HUT 102 and CTCL-3, and in fresh peripheral blood lymphocytes obtained from a patient with a cutaneous T-cell lymphoma (mycosis fungoides). The cell lines continuously produce these viruses, which are collectively referred to as HTLV, strain CR (HTLV[CR]). Originally, the production of virus from HUT 102 cells required induction with 5-iodo-2'-deoxyuridine, but the cell line became a constitutive producer of virus at its 56th passage. Cell line CTCL-3 has been a constitutive producer of virus from its second passage in culture. Both mature and immature extracellular virus particles were seen in thin-section electron micrographs of fixed, pelleted cellular material; on occasion, typical type C budding virus particles were seen. No form of intracellular virus particle has been seen. Mature particles were 100-110 nm in diameter, consisted of an electron-dense core surrounded by an outer membrane separated by an electron-lucent region, banded at a density of 1.16 g/ml on a continuous 25-65% sucrose gradient, and contained 70S RNA and a DNA polymerase activity typical of viral reverse transcriptase (RT;RNA-dependent DNA nucleotidyltransferase). Under certain conditions of assay, HTLV[CR] RT showed cation preference for Mg 2+ over Mn 2+, distinct from the characteristics of cellular DNA polymerases purified from human lymphocytes and the RT from most type C viruses. Antibodies to cellular DNA polymerase γ and antibodies against RT purified from several animal retroviruses failed to detectably interact with HTLV[CR]RT under conditions that were positive for the respective homologous DNA polymerase, demonstrating a lack of close relationship of HTLV[CR]RT to cellular DNA polymerases γ or RT of these viruses. Six major proteins, with sizes of approximately 10,000, 13,000, 19,000, 24,000, 42,000 and 52,000 daltons were apparent when doubly banded, disrupted HTLV[CR] particles were chromatographed on a NaDodSO 4/polyacrylamide gel. The number of these particle-associated proteins is consistent with the expected proteins of a retrovirus, but the sizes of some are distinct from those of most known retroviruses of the primate subgroups.

AB - Retrovirus particles with type C morphology were found in two T-cell lymphoblastoid cell lines, HUT 102 and CTCL-3, and in fresh peripheral blood lymphocytes obtained from a patient with a cutaneous T-cell lymphoma (mycosis fungoides). The cell lines continuously produce these viruses, which are collectively referred to as HTLV, strain CR (HTLV[CR]). Originally, the production of virus from HUT 102 cells required induction with 5-iodo-2'-deoxyuridine, but the cell line became a constitutive producer of virus at its 56th passage. Cell line CTCL-3 has been a constitutive producer of virus from its second passage in culture. Both mature and immature extracellular virus particles were seen in thin-section electron micrographs of fixed, pelleted cellular material; on occasion, typical type C budding virus particles were seen. No form of intracellular virus particle has been seen. Mature particles were 100-110 nm in diameter, consisted of an electron-dense core surrounded by an outer membrane separated by an electron-lucent region, banded at a density of 1.16 g/ml on a continuous 25-65% sucrose gradient, and contained 70S RNA and a DNA polymerase activity typical of viral reverse transcriptase (RT;RNA-dependent DNA nucleotidyltransferase). Under certain conditions of assay, HTLV[CR] RT showed cation preference for Mg 2+ over Mn 2+, distinct from the characteristics of cellular DNA polymerases purified from human lymphocytes and the RT from most type C viruses. Antibodies to cellular DNA polymerase γ and antibodies against RT purified from several animal retroviruses failed to detectably interact with HTLV[CR]RT under conditions that were positive for the respective homologous DNA polymerase, demonstrating a lack of close relationship of HTLV[CR]RT to cellular DNA polymerases γ or RT of these viruses. Six major proteins, with sizes of approximately 10,000, 13,000, 19,000, 24,000, 42,000 and 52,000 daltons were apparent when doubly banded, disrupted HTLV[CR] particles were chromatographed on a NaDodSO 4/polyacrylamide gel. The number of these particle-associated proteins is consistent with the expected proteins of a retrovirus, but the sizes of some are distinct from those of most known retroviruses of the primate subgroups.

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U2 - 10.1073/pnas.77.12.7415

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