Mitogen requirements for the in vitro propagation of cutaneous T-cell lymphomas

A. F. Gazdar, D. N. Carney, P. A. Bunn, E. K. Russell, E. S. Jaffe, G. P. Schechter, J. G. Guccion

Research output: Contribution to journalArticle

320 Citations (Scopus)

Abstract

The authors attempted to generate continuous in vitro cultures from patients with mycosis fungoides and the Sezary syndrome (cutaneous T-cell lymphomas, CTCL). Using conventional culture techniques without mitogen stimulation, multiple attempts (32 specimens from 25 patients) failed to replicate T cells, but 6 B-lymphoblastoid cultures were established. Athymic nude mice injected by a variety of routes with CTCL cells from 13 patients failed to develop tumors; however, the B-lymphoblastoid cultures were tumorigenic. Lymphocytes from 6 healthy donors and CTCL cells from 7 patients were seeded in growth medium supplemented with one of the following mitogens: phytohemagglutinin (PHA), concanavalin A (Con-A), lymphocyte-conditioned medium (LyCM), pokeweed mitogen (PWM), and staphylococcal protein-A. Normal lymphocytes failed to replicate for more than a few days except in LyCM, where viability and replication remained absolutely dependent on the continued presence of the mitogen. In contrast, mitogen-induced proliferation of CTCL cells was variable, and 1 or more of 4 mitogens induced replication in 4 CTCL cultures. There was partial correlation between the ability of mitogens to stimulate CTCL cells in lymphocyte transformation assays and their ability to act as growth factors. CTCL cells were more resistant to the toxic effects of mitogen than lymphocytes from normal donors, permitting the use of mitogens as long-term growth factors. Two long-term CTCL cultures were established, one after stimulation with Con-A, and the other with LyCM. The cultures required mitogens only for initial growth and have proliferated without mitogens for over a year. The cultures retained many of the features of the CTCL cells present in the patients from whom they were derived, but differed from each other in morphology, E-rosette formation, DNA content, and tumorigenicity. Cytochemical studies provided further evidence of their T-cell origin. Both cultures released macrophage inhibitory factor.

Original languageEnglish (US)
Pages (from-to)409-417
Number of pages9
JournalBlood
Volume55
Issue number3
StatePublished - 1980

Fingerprint

Cutaneous T-Cell Lymphoma
T-cells
Mitogens
Lymphocytes
Cell culture
Conditioned Culture Medium
Concanavalin A
Nude Mice
Intercellular Signaling Peptides and Proteins
Cell Culture Techniques
In Vitro Techniques
Tissue Donors
Sezary Syndrome
T-Lymphocytes
Rosette Formation
Pokeweed Mitogens
Culture Techniques
Mycosis Fungoides
Poisons
Staphylococcal Protein A

ASJC Scopus subject areas

  • Hematology

Cite this

Gazdar, A. F., Carney, D. N., Bunn, P. A., Russell, E. K., Jaffe, E. S., Schechter, G. P., & Guccion, J. G. (1980). Mitogen requirements for the in vitro propagation of cutaneous T-cell lymphomas. Blood, 55(3), 409-417.

Mitogen requirements for the in vitro propagation of cutaneous T-cell lymphomas. / Gazdar, A. F.; Carney, D. N.; Bunn, P. A.; Russell, E. K.; Jaffe, E. S.; Schechter, G. P.; Guccion, J. G.

In: Blood, Vol. 55, No. 3, 1980, p. 409-417.

Research output: Contribution to journalArticle

Gazdar, AF, Carney, DN, Bunn, PA, Russell, EK, Jaffe, ES, Schechter, GP & Guccion, JG 1980, 'Mitogen requirements for the in vitro propagation of cutaneous T-cell lymphomas', Blood, vol. 55, no. 3, pp. 409-417.
Gazdar AF, Carney DN, Bunn PA, Russell EK, Jaffe ES, Schechter GP et al. Mitogen requirements for the in vitro propagation of cutaneous T-cell lymphomas. Blood. 1980;55(3):409-417.
Gazdar, A. F. ; Carney, D. N. ; Bunn, P. A. ; Russell, E. K. ; Jaffe, E. S. ; Schechter, G. P. ; Guccion, J. G. / Mitogen requirements for the in vitro propagation of cutaneous T-cell lymphomas. In: Blood. 1980 ; Vol. 55, No. 3. pp. 409-417.
@article{5f99fe298630451aa37e5e9d042c5d0a,
title = "Mitogen requirements for the in vitro propagation of cutaneous T-cell lymphomas",
abstract = "The authors attempted to generate continuous in vitro cultures from patients with mycosis fungoides and the Sezary syndrome (cutaneous T-cell lymphomas, CTCL). Using conventional culture techniques without mitogen stimulation, multiple attempts (32 specimens from 25 patients) failed to replicate T cells, but 6 B-lymphoblastoid cultures were established. Athymic nude mice injected by a variety of routes with CTCL cells from 13 patients failed to develop tumors; however, the B-lymphoblastoid cultures were tumorigenic. Lymphocytes from 6 healthy donors and CTCL cells from 7 patients were seeded in growth medium supplemented with one of the following mitogens: phytohemagglutinin (PHA), concanavalin A (Con-A), lymphocyte-conditioned medium (LyCM), pokeweed mitogen (PWM), and staphylococcal protein-A. Normal lymphocytes failed to replicate for more than a few days except in LyCM, where viability and replication remained absolutely dependent on the continued presence of the mitogen. In contrast, mitogen-induced proliferation of CTCL cells was variable, and 1 or more of 4 mitogens induced replication in 4 CTCL cultures. There was partial correlation between the ability of mitogens to stimulate CTCL cells in lymphocyte transformation assays and their ability to act as growth factors. CTCL cells were more resistant to the toxic effects of mitogen than lymphocytes from normal donors, permitting the use of mitogens as long-term growth factors. Two long-term CTCL cultures were established, one after stimulation with Con-A, and the other with LyCM. The cultures required mitogens only for initial growth and have proliferated without mitogens for over a year. The cultures retained many of the features of the CTCL cells present in the patients from whom they were derived, but differed from each other in morphology, E-rosette formation, DNA content, and tumorigenicity. Cytochemical studies provided further evidence of their T-cell origin. Both cultures released macrophage inhibitory factor.",
author = "Gazdar, {A. F.} and Carney, {D. N.} and Bunn, {P. A.} and Russell, {E. K.} and Jaffe, {E. S.} and Schechter, {G. P.} and Guccion, {J. G.}",
year = "1980",
language = "English (US)",
volume = "55",
pages = "409--417",
journal = "Blood",
issn = "0006-4971",
publisher = "American Society of Hematology",
number = "3",

}

TY - JOUR

T1 - Mitogen requirements for the in vitro propagation of cutaneous T-cell lymphomas

AU - Gazdar, A. F.

AU - Carney, D. N.

AU - Bunn, P. A.

AU - Russell, E. K.

AU - Jaffe, E. S.

AU - Schechter, G. P.

AU - Guccion, J. G.

PY - 1980

Y1 - 1980

N2 - The authors attempted to generate continuous in vitro cultures from patients with mycosis fungoides and the Sezary syndrome (cutaneous T-cell lymphomas, CTCL). Using conventional culture techniques without mitogen stimulation, multiple attempts (32 specimens from 25 patients) failed to replicate T cells, but 6 B-lymphoblastoid cultures were established. Athymic nude mice injected by a variety of routes with CTCL cells from 13 patients failed to develop tumors; however, the B-lymphoblastoid cultures were tumorigenic. Lymphocytes from 6 healthy donors and CTCL cells from 7 patients were seeded in growth medium supplemented with one of the following mitogens: phytohemagglutinin (PHA), concanavalin A (Con-A), lymphocyte-conditioned medium (LyCM), pokeweed mitogen (PWM), and staphylococcal protein-A. Normal lymphocytes failed to replicate for more than a few days except in LyCM, where viability and replication remained absolutely dependent on the continued presence of the mitogen. In contrast, mitogen-induced proliferation of CTCL cells was variable, and 1 or more of 4 mitogens induced replication in 4 CTCL cultures. There was partial correlation between the ability of mitogens to stimulate CTCL cells in lymphocyte transformation assays and their ability to act as growth factors. CTCL cells were more resistant to the toxic effects of mitogen than lymphocytes from normal donors, permitting the use of mitogens as long-term growth factors. Two long-term CTCL cultures were established, one after stimulation with Con-A, and the other with LyCM. The cultures required mitogens only for initial growth and have proliferated without mitogens for over a year. The cultures retained many of the features of the CTCL cells present in the patients from whom they were derived, but differed from each other in morphology, E-rosette formation, DNA content, and tumorigenicity. Cytochemical studies provided further evidence of their T-cell origin. Both cultures released macrophage inhibitory factor.

AB - The authors attempted to generate continuous in vitro cultures from patients with mycosis fungoides and the Sezary syndrome (cutaneous T-cell lymphomas, CTCL). Using conventional culture techniques without mitogen stimulation, multiple attempts (32 specimens from 25 patients) failed to replicate T cells, but 6 B-lymphoblastoid cultures were established. Athymic nude mice injected by a variety of routes with CTCL cells from 13 patients failed to develop tumors; however, the B-lymphoblastoid cultures were tumorigenic. Lymphocytes from 6 healthy donors and CTCL cells from 7 patients were seeded in growth medium supplemented with one of the following mitogens: phytohemagglutinin (PHA), concanavalin A (Con-A), lymphocyte-conditioned medium (LyCM), pokeweed mitogen (PWM), and staphylococcal protein-A. Normal lymphocytes failed to replicate for more than a few days except in LyCM, where viability and replication remained absolutely dependent on the continued presence of the mitogen. In contrast, mitogen-induced proliferation of CTCL cells was variable, and 1 or more of 4 mitogens induced replication in 4 CTCL cultures. There was partial correlation between the ability of mitogens to stimulate CTCL cells in lymphocyte transformation assays and their ability to act as growth factors. CTCL cells were more resistant to the toxic effects of mitogen than lymphocytes from normal donors, permitting the use of mitogens as long-term growth factors. Two long-term CTCL cultures were established, one after stimulation with Con-A, and the other with LyCM. The cultures required mitogens only for initial growth and have proliferated without mitogens for over a year. The cultures retained many of the features of the CTCL cells present in the patients from whom they were derived, but differed from each other in morphology, E-rosette formation, DNA content, and tumorigenicity. Cytochemical studies provided further evidence of their T-cell origin. Both cultures released macrophage inhibitory factor.

UR - http://www.scopus.com/inward/record.url?scp=0018903124&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0018903124&partnerID=8YFLogxK

M3 - Article

C2 - 6244013

AN - SCOPUS:0018903124

VL - 55

SP - 409

EP - 417

JO - Blood

JF - Blood

SN - 0006-4971

IS - 3

ER -