T Cell Growth Factor from Human Splenic Cell Cultures: Conditions for Its Production, and Its Utilization for Maintenance of Cytotoxic T Cell Lines

Tanaka, Yuetsu; Sugamura, Kazuo; Hinuma, Yorio

doi:10.1111/j.1348-0421.1981.tb00114.x

Cited by 15 publications

(7 citation statements)

References 15 publications

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“…The radioactivities bound to cells were measured. ATLSA, cell surface antigens reacting with anti-HTLV-positive human serum of an ATL patient, were also assayed by RIA using 1251-labelled protein A HTLV-bearing T-cell lines were established from mouse Ig or protein A was calculated as follows: PBL of four ATL patients and five healthy HTLVcarriers with medium containing IL2 as described precounts in test sample ly, limiting numbers of PBL of ATL patients were counts in control sample cultured in RPMI1640 medium supplement with 20 % Control samples were 0.5 % BSA in PBS used for Fcs and 25% crude IL2 which were Prepared from dilution of antibodies in assays of surface markers and human spleen-cell cultures (Tanaka et al, 1981). Four anti-HTLV-negative human Semm in assays for IL2-dependent cell lines thus obtained from the ATL ATLSA.…”

Section: Radioimmunoassay (Ria)mentioning

confidence: 99%

Cell surface phenotypes and expression of viral antigens of various human cell lines carrying human T‐cell leukemia virus

Sugamura

Fujii

Kannagi

et al. 1984

Intl Journal of Cancer

Self Cite

183

106

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Human T-cell leukemia/lymphoma virus (HTLV)-carrying cells from various origins were characterized by cell surface markers and expression of HTLV antigens. Eight cell lines named TCL were obtained by transformation of peripheral blood leukocytes (PBL) of healthy donors or HTLV carriers in cocultures with HTLV-producer MT-2 cells. Nine cell lines named ILT were interleukin 2 (IL2)-dependent cell lines cloned from PBL of ATL patients and healthy HTLV-carriers. Tc-Kan9 cell line was also an IL2-dependent cell line clonally established from PBL culture stimulated with autologous TCL cells. Five cell lines named TL were established in vitro directly from PBL of an adult T-cell leukemia (ATL) patient and from ILT cells of an ATL patient and three HTLV-carriers, respectively, to grow autonomously without IL2. All the TCLs, ILTs, TLs and Tc-Kan9 possessed Leu-I antigen, a pan-T-cell marker. Leu3a antigen, a helper/inducer T-cell marker, was expressed on five of eight TCLs and all of the ILTs and TLs. Leu-2a, a cytotoxic/suppressor T-cell marker, was detected only on Tc-Kan9 but not others. Fresh ATL leukemic cells of patients had a helper/inducer T-cell marker. Ia, OKT9 and Tac antigens, markers for activated and differentiated T cells, were strongly expressed on all of the cell lines tested and fresh ATL leukemic cells were weakly positive for these antigens. Expression of HTLV antigens detected by mouse monoclonal antibodies and an ATL-patient serum varied among these cell lines. One TL, two ILTs and most of the fresh ATL leukemic cells did not express HTLV antigens on the cell surface. The other cell lines were all positive for the surface viral antigens. However, molecular species of antigens defined by radioimmunoprecipitation with an ATL-patient serum were not always identical among the cell lines. Molecular weights of polypeptides detectable in most of the cell lines were 62K, 46K, 40K, 24K, 21K and 19K which could never be detected in several control T-cell lines. 68K and 28K polypeptides were frequently detected in MT-2 and TGLs. GIN14, a mouse monoclonal antibody against HTLV core protein (p19) detected not only p19 in various cell lines but also p28, p29, p31 or p40 in certain cell lines tested. B-cell lines named LCL were established and cloned from PBL of two HTLV-carriers by EB-virus-induced transformation and they also expressed HTLV antigens, Ia, OKT9 and Tac antigens.(ABSTRACT TRUNCATED AT 400 WORDS)

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Section: Radioimmunoassay (Ria)mentioning

confidence: 99%

Cell surface phenotypes and expression of viral antigens of various human cell lines carrying human T‐cell leukemia virus

Sugamura

Fujii

Kannagi

et al. 1984

Intl Journal of Cancer

Self Cite

183

106

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“…Details of the method for TCGF preparation were described elsewhere (8 (Table 3). On primary cloning at 1,000 cells per well, about eight ATLA-positive clones were obtained in 288 wells.…”

mentioning

confidence: 99%

Healthy carriers of a human retrovirus, adult T-cell leukemia virus (ATLV): demonstration by clonal culture of ATLV-carrying T cells from peripheral blood.

Gotoh¹,

Sugamura²,

Hinuma³

1982

Proc. Natl. Acad. Sci. U.S.A.

113

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Adult T-cell leukemia virus (ATLV) or ATLV-associated antigen (ATLA)-positive cell clones were isolated from peripheral blood lymphocytes of all five anti-ATLA-seropositive healthy adults tested by a limiting-dilution culture method in the presence of T-cell growth factor (TCGF) but from none of six seronegative adults similarly tested. Although ATLA-positive cells were not always detected in mass cultures of the seropositive lym-' phocytes, they were found consistently in T-cell cloned cultures of those lymphocytes. Continuous cultures of the ATLA-positive clones obtained were dependent on TCGF. Five clones derived from each of the five donors were maintained for >4 months and, during culture, all of them acquired the ability to grow without added TCGF. The ATLA-positive clones formed rosettes with erythrocytes and expressed Leu 1, Leu 3a, and Leu 4 antigens but not Leu 2a antigen. These results indicate that anti-ATLA-seropositive healthy individuals carry ATLV in T cells circulating in their peripheral blood.The human leukemia adult T-cell leukemia (ATL) has been recognized to be endemic in restricted areas of Japan (1-3). Recently, ATL virus (ATLV) was detected in ATL-derived human T-cell lines (4, 5) and characterized as a retrovirus (6), and a close relationship between ATL and ATLV was suggested by serologic and biochemical analyses. In these studies, antibodies to antigens associated.with ATLV-producer cell lines (ATLA), which were proved to be specific for ATLV (ref. 6

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“…The nonhuman primate cell lines listed in Table 5 were established from peripheral blood lymphocytes (PBL) co-cultivated with irradiated MT-2 cells or 2CM cells, which are T-cell line cells derived from cynomolgus monkey PBL co-cultivated with MT-2 cells (22). All cell lines were cultivated in RPMI-1640 medium supplemented with 16% fetal calf serum (FCS), 100 IU of penicillin per ml and 100 pg of streptomycin per ml in the presence or absence of 25% crude human splenic IL-2 preparation (20). For some experiments, PBL samples from humans and various nonhuman primates were activated with 1% phytohemagglutinin-M (PHA) (Difco) and cultured with crude human IL-2 for 3 to 14 days, and the recovered cells were used.…”

Section: Methodsmentioning

confidence: 99%

New Monoclonal Antibodies That Define Multiple Epitopes and a Human‐Specific Marker on the Interleukin 2 Receptor Molecules of Primates

Tanaka

Inoi

Tozawa

et al. 1986

Microbiology and Immunology

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Twelve hybridoma cell lines producing monoclonal antibodies to the human interleukin 2 (IL-2) receptor (IL-2R) molecule were prepared. These antibodies were characterized by competitive antibody-binding assay and sequential immunoprecipitation assay with four known monoclonal antibodies to the human IL-2R molecule. The twelve new monoclonal antibodies were divided among the four known antibody types, the HIEI-, H-A26-, H-31-, and anti-Tac-type, and an additional new type, the H-48-type. The H-48 antibody did not compete with any other antibodies in the competitive binding assay. The binding of 1-25I-IL-2 to MT-2 cells and the IL-2-dependent growth of normal activated T-cells were both strongly inhibited by all the H-31-and anti-Tac-type antibodies, and partially or slightly inhibited by HIEI-and H-A26-type antibodies, but were not inhibited by the H-48 antibody. Thus, the same type of monoclonal antibodies had a similar effect on the function of IL-2R. These results suggest that epitopes for the same type of antibodies could be single identical epitopes or epitopes closely associated with each other. On the other hand, these antibodies also reacted variously with a panel of various human and simian lymphoid cell lines immortalized with human T-cell leukemia virus type-I (HTLV-I) : the H-45 antibody reacted only with the human cell lines, the H-Cl and H-44 and H-47 antibodies reacted with human and ape cell lines, and the other antibodies reacted with cell lines of humans, apes and Old and New World monkeys. These differences in the reactivity of the antibodies with the primate cell lines suggest that the antigenic structure of the IL-2R molecule changed during evolutionary divergence of the primates.

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T Cell Growth Factor from Human Splenic Cell Cultures: Conditions for Its Production, and Its Utilization for Maintenance of Cytotoxic T Cell Lines

Abstract: We prepared the T

Cited by 15 publications

References 15 publications

Cell surface phenotypes and expression of viral antigens of various human cell lines carrying human T‐cell leukemia virus

Cell surface phenotypes and expression of viral antigens of various human cell lines carrying human T‐cell leukemia virus

Healthy carriers of a human retrovirus, adult T-cell leukemia virus (ATLV): demonstration by clonal culture of ATLV-carrying T cells from peripheral blood.

New Monoclonal Antibodies That Define Multiple Epitopes and a Human‐Specific Marker on the Interleukin 2 Receptor Molecules of Primates

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