RESUMO
Human T cell hybrids were generated by fusing lectin-activated normal and leukemic human T cells with an aminopterin-sensitive human T cell line. This mutant cell line, designated CEM-T15, was derived from the human T cell line CEM after chemical mutagenesis with ethane methylsulfonate and subsequent culture in medium containing 6-thioguanine. After polyethylene glycol-induced fusion, the cells were cultured in hypoxanthine-aminopterin-thymidine selective medium. More than 5 wk after fusion, evidence for successful hybridization was obtained by three independent criteria: (a) The majority of the cultures contained cells expressing the OKT3 surface antigen: this antigen is expressed on normal T cells but not on CEM-T15 cells. (b) Most of the cultures contained polyploid cells. (c) Some of the cultures provided helper activity in the generation of antibody-forming cells. This functional activity is absent from the CEM-T15 parental cell line. Evidence for functional stability of the hybrids greater than 20 wk after fusion was provided by several clones that not only continue growing exponentially but also maintain expression of OKT3 surface antigen and high levels of helper function. These T cell hybrids constructed using antigen-specific human T cells should be of considerable importance in further studies of the immunobiology of human T cells.
Assuntos
Células Híbridas/citologia , Linfócitos T/citologia , Aminopterina/farmacologia , Animais , Diferenciação Celular , Linhagem Celular , Células Clonais/imunologia , Meios de Cultura , Humanos , Células Híbridas/imunologia , Cariotipagem , Ativação Linfocitária , Coelhos , Linfócitos T/imunologia , Tioguanina/farmacologiaRESUMO
In this report, we explored the functional heterogeneity within the OKT4+ subset of human T cells. Evidence was obtained that although in vitro pokeweed mitogen-activated OKT4+ cells can function as radioresistant helper cells, these activated OKT4+ cells could also exert potent feedback suppression. Despite the induction of suppressor cells after pokeweed mitogen activation, the OKT4+ population maintains its original OKT3+, OKT4+, nd OKT8- surface phenotype. The suppressor cells contained within the activated OKT4+ population were found to be radiosensitive. Importantly, the suppression mediated by activated OKT4+ cells required the presence of radiosensitive cells contained within the resting OKT4+ population. Taken together, these results suggest that the OKT4+ subset of human T cells contains cells that can be activated to differentiate into suppressor cells independent of OKT8+ cells.
Assuntos
Anticorpos/análise , Linfócitos T Reguladores/imunologia , Linfócitos T/classificação , Anticorpos Monoclonais , Linfócitos B/citologia , Linfócitos B/efeitos da radiação , Diferenciação Celular , Humanos , FenótipoRESUMO
CASE REPORT: We report the case of a 10-year-old boy with a progressive bilateral vitelliform macular dystrophy, and his father with terminal stage disease in both eyes; we studied the development and progression of this condition with optical coherence tomography. DISCUSSION: Optical coherence tomography is a useful noninvasive tool that complements other diagnostic modalities and improves the follow up assessment. It provides additional information on the morphology of the lesion as well as identifying secondary changes in the adjacent retina. It also demonstrates the location of any yellowish material under the sensory retina.
Assuntos
Degeneração Macular/diagnóstico , Tomografia de Coerência Óptica , Criança , Progressão da Doença , Angiofluoresceinografia , Fundo de Olho , Humanos , Degeneração Macular/classificação , Degeneração Macular/genética , Masculino , Pessoa de Meia-Idade , Acuidade VisualRESUMO
Two murine monoclonal antibodies (BE1 and BE2), produced by using leukemic helper T cells from a patient with cutaneous T-cell lymphoma (CTCL) as immunogens, reacted selectively with CTCL lymphocytes and some transformed cultured lymphocytes, as determined by radioimmunoassay (RIA) and indirect immunofluorescence (IIF). BE1 reacted significantly (P less than or equal to 0.001) with leukemic CTCL lymphocytes and with CTCL cells from infiltrated lymph nodes (RIA, mean +/- SD = 776 +/- 275 cpm), as compared with background counts (263 +/- 68). BE1 binding to normal blood mononuclear cells (RIA, mean +/- SD = 283 +/- 58 cpm) was indistinguishable from background. BE1 also reacted with Epstein-Barr virus (EBV)-transformed B-cell lines (RIA, mean +/- SD = 794 +/- 230) and some long-term T-cell lines. BE1 did not react with the majority of lymphoid cell lines or tumor cell lines tested. BE1 also did not react with any normal tissues screened by IIF. BE1 precipitated a molecule from CTCL cells that, under reducing conditions, has two components with molecular mass of 27,200 and 25,800 D. BE2 also reacted significantly (P less than or equal to 0.001) with CTCL cells from two of four patients (RIA, mean +/- SD = 519 +/- 113 cpm). The binding of BE2 to normal mononuclear cells was indistinguishable from background (309 +/- 38 cpm). BE2 also reacted with an antigen present on EBV-B-cell lines (RIA, mean +/- SD = 654 +/- 194) and MOLT 3 and HUT 78 T-cell lines. BE2 reacted with an antigen expressed on a subpopulation of lymphocytes from five of eight patients with B-cell CLL studied by IIF (mean +/- SD = 18 +/- 6). Other long-term T-cell lines and tumor cell lines studied by IIF were unreactive with BE2. BE2 did not react with any of the normal tissues studied. BE2 precipitated a molecule (78,000 D) from CTCL cells and EBV-B cells with a single component under reducing conditions. Immunoperoxidase-labeled BE1 and BE2 reacted with CTCL cells in frozen sections of infiltrated lymph nodes and skin. In addition, BE1 and BE2 reacted with blood lymphocytes from 16 of 21 patients whose CTCL had otherwise been considered localized to skin. These two monoclonal antibodies react with tumor antigens associated with CTCL and appear to be useful in the diagnosis of this disorder.
Assuntos
Anticorpos Monoclonais/imunologia , Antígenos de Neoplasias/análise , Linfoma/diagnóstico , Neoplasias Cutâneas/diagnóstico , Linfócitos T Auxiliares-Indutores/imunologia , Especificidade de Anticorpos , Linhagem Celular , Humanos , Linfoma/imunologia , Neoplasias Cutâneas/imunologiaRESUMO
Resisting death is a central hallmark of cancer cells. Tumors rely on a number of genetic mechanisms to avoid apoptosis, and alterations in mRNA alternative splicing are increasingly recognized to have a role in tumorigenesis. In this study, we identify the splicing regulator SLU7 as an essential factor for the preservation of hepatocellular carcinoma (HCC) cells viability. Compared with hepatocytes, SLU7 expression is reduced in HCC cells; however, further SLU7 depletion triggered autophagy-related cellular apoptosis in association with the overproduction of reactive oxygen species. Remarkably, these responses were not observed in primary human hepatocytes or in the well-differentiated HepaRG cell line. Mechanistically, we demonstrate that SLU7 binds the C13orf25 primary transcript in which the polycistronic oncomir miR-17-92 cluster is encompassed, and is necessary for its processing and expression. SLU7 knockdown altered the splicing of the C13orf25 primary transcript, and markedly reduced the expression of its miR-17, miR-20 and miR-92a constituents. This led to the upregulation of CDKN1A (P21) and BCL2L11 (BIM) expression, two bona fide targets of the miR-17-92 cluster and recognized mediators of its pro-survival and tumorigenic activity. Interestingly, altered splicing of miR-17-92 and downregulation of miR-17 and miR-20 were not observed upon SLU7 knockdown in non-transformed hepatocytes, but was found in other (HeLa, H358) but not in all (Caco2) non-hepatic tumor cells. The functional relevance of miR-17-92 dysregulation upon SLU7 knockdown was established when oxidative stress, autophagy and apoptosis were reversed by co-transfection of HCC cells with a miR-17 mimic. Together, these findings indicate that SLU7 is co-opted by HCC cells and other tumor cell types to maintain survival, and identify this splicing regulator as a new determinant for the expression of the oncogenic miR-17-92 cluster. This novel mechanism may be exploited for the development of antitumoral strategies in cancers displaying such SLU7-miR-17-92 crosstalk.
Assuntos
Processamento Alternativo/genética , Carcinoma Hepatocelular/genética , Neoplasias Hepáticas/genética , MicroRNAs/genética , Fatores de Processamento de RNA/genética , Apoptose/genética , Autofagia/genética , Células CACO-2 , Carcinogênese/genética , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Sobrevivência Celular/genética , Regulação Neoplásica da Expressão Gênica , Células Hep G2 , Hepatócitos/metabolismo , Hepatócitos/patologia , Humanos , Neoplasias Hepáticas/patologia , RNA Longo não CodificanteRESUMO
CASE REPORT: The case is reported of acute retinal necrosis with bilateral involvement due to Varicella Zoster virus in a 77 year-old man. Polymerase chain reaction (PCR) of aqueous humor was positive for Varicella Zoster virus (VZV). He developed a Kyrieleis' vasculitis a month after the starting treatment, when the PCR analysis was negative. DISCUSSION: PCR is a quick and safe technique, with a high sensitivity and specificity of 97%, useful to diagnose and monitor the viral activity. The intervention must be urgent, due to the dramatically rapid evolution. Oral famciclovir oral is good alternative owing to its bioavailability.
Assuntos
Herpes Zoster Oftálmico/complicações , Síndrome de Necrose Retiniana Aguda/etiologia , Idoso , Humor Aquoso/virologia , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/imunologia , Herpesvirus Humano 3/genética , Herpesvirus Humano 3/isolamento & purificação , Humanos , Hospedeiro Imunocomprometido , Masculino , Reação em Cadeia da Polimerase , Neoplasias da Próstata/complicações , Neoplasias da Próstata/imunologia , Vasculite Retiniana/etiologia , Lâmpada de FendaRESUMO
Human T-cell hybrids with helper activity were obtained after fusion of phytohemagglutinin-activated normal human T cells with a 6-thioguanine-resistant, aminopterin-sensitive human T-cell line. This mutant line, designated CEM-T15, was derived from the human T-cell line CEM after mutagenesis with ethyl methanesulfonate. The polyethylene glycol induced fusion and the selection in hypoxanthine- aminopeterin -thymidine medium were performed by modification of standard somatic cell hybridization techniques. After fusion, the strategy for selecting hybrids consisted in screening growing cultures for the presence of cells expressing the OKT3 cell surface differentiation antigen. OKT3 was chosen because it is present in 85-95% of normal human T cells but absent from CEM-T15 cells. Thus, OKT3+ cells growing 5-7 weeks after fusion most likely represented hybrids between normal T cells (OKT3+) and continuously growing CEM-T15 cells (OKT3-). Several of the hybrids were tested for their capacity to promote pokeweed mitogen-induced antibody production by B cells. These experiments demonstrated that many of the hybrids had helper activity. Periodical testing of these uncloned hybrids for helper activity revealed functional instability, with most of the hybrids losing helper activity after 20 weeks of continuous culture. However, early and repeated cloning of the same hybrids resulted in a series of hybrid clones with helper activity still present more than 8 months after fusion. In more recent fusions, we have demonstrated that human helper hybrids producing helper factor(s) can also be obtained. These and similar hybrids with different functions will be of considerable importance in further studies of the immunobiology of human T lymphocytes.
Assuntos
Células Híbridas/fisiologia , Linfócitos T Auxiliares-Indutores/imunologia , Linfócitos T/fisiologia , Antígenos de Superfície/análise , Bromodesoxiuridina/toxicidade , Linhagem Celular , Cromossomos Humanos/fisiologia , Células Clonais , Técnicas de Cultura/métodos , Resistência a Medicamentos , Humanos , Células Híbridas/imunologia , Leucemia Linfoide , Ativação Linfocitária , Mutação , Linfócitos T/imunologia , Tioguanina/toxicidadeRESUMO
CASE REPORT: A 25-year-old woman noticed a painless yellow-orange mass on her right eye. Her visual acuity was 20/20 in both eyes, and a slit-lamp examination showed a yellow-orange mass located at the superior limbus of the right eye. No other ocular abnormalities were observed. DISCUSSION: Surgical excision was carried out and the lesion was sent for histological examination. This showed a granulomatous lesion, rich in Touton-type giant cells, features that are strongly suggestive of juvenile xanthogranuloma (JXG). Ocular involvement occurs in 10% of cases of JXG.
Assuntos
Limbo da Córnea/patologia , Xantogranuloma Juvenil/diagnóstico , Adulto , Idade de Início , Diagnóstico Diferencial , Feminino , Células Gigantes/patologia , Histiócitos/patologia , Humanos , Limbo da Córnea/cirurgia , Xantogranuloma Juvenil/epidemiologia , Xantogranuloma Juvenil/patologia , Xantogranuloma Juvenil/cirurgiaRESUMO
No disponible
Assuntos
Humanos , Masculino , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Técnicas de Apoio para a Decisão , Sistemas de Manutenção da Vida/instrumentação , Unidades de Terapia Intensiva , Estudos Prospectivos , Intervalos de Confiança , Estatísticas não ParamétricasRESUMO
No disponible
Assuntos
Humanos , Adulto , Estudos Soroepidemiológicos , Doadores de Tecidos/estatística & dados numéricos , Anticorpos Anti-HTLV-I/análise , Anticorpos Anti-HTLV-II/análise , Infecções por HTLV-I/epidemiologia , Infecções por HTLV-II/epidemiologia , Anticorpos Anti-HTLV-I/imunologia , Anticorpos Anti-HTLV-II/imunologia , Vírus Linfotrópico T Tipo 1 Humano/imunologia , Vírus Linfotrópico T Tipo 2 Humano/imunologiaRESUMO
CASO CLÍNICO: Presentamos a un paciente de 77 años, con un cuadro de necrosis retiniana aguda con afectación bilateral; la reacción en cadena de la polimerasa (PCR) de la muestra de humor acuoso fue positiva al virus de varicela zóster. En su evolución desarrolla vasculitis de Kyrieleis al mes de inicio del tratamiento y con el análisis por PCR negativo. DISCUSIÓN: La PCR es un método rápido, con una sensibilidad y especificidad del 97%. La actuación debe ser urgente por la rapidez de la progresión. El famciclovir oral es buena alternativa por su mejor biodisponibilidad
CASE REPORT: The case is reported of acute retinal necrosis with bilateral involvement due to Varicella Zoster virus in a 77 year-old man. Polymerase chain reaction (PCR) of aqueous humor was positive for Varicella Zoster virus (VZV). He developed a Kyrieleis vasculitis a month after the starting treatment, when the PCR analysis was negative. DISCUSSION: PCR is a quick and safe technique, with a high sensitivity and specificity of 97%, useful to diagnose and monitor the viral activity. The intervention must be urgent, due to the dramatically rapid evolution. Oral famciclovir oral is good alternative owing to its bioavailability
Assuntos
Adulto , Humanos , Masculino , Síndrome de Necrose Retiniana Aguda/metabolismo , Síndrome de Necrose Retiniana Aguda/patologia , Herpes Zoster/patologia , Retinite/genética , Vasculite/diagnóstico , Reação em Cadeia da Polimerase/métodos , Síndrome de Necrose Retiniana Aguda/congênito , Síndrome de Necrose Retiniana Aguda/genética , Herpes Zoster/diagnóstico , Retinite/metabolismo , Vasculite/complicações , Reação em Cadeia da PolimeraseRESUMO
Human T lymphocytes bearing the cell surface antigen T4 are functionally heterogeneous, exerting helper/inducer, suppressor-inducer, suppressor-effector, and cytotoxic activities. Other cell surface antigens with a more restricted expression may help separate T4+ lymphocytes into functionally distinct subsets. This report describes the regulatory functions of T4+ lymphocytes fractionated by the monoclonal antibody 5/9, which detects a cell surface antigen present on 50-60% of T4+ lymphocytes. The results indicate that both 5/9+ and 5/9- T4 subsets contain helper/inducer and suppressor-inducer cells. Suppressor-effector activity, however, is found predominantly within the 5/9+ T4 subset. The 5/9 antibody thus identifies the suppressor-effector subset of T4+ lymphocytes, although it does not distinguish between T4+ cells with or without helper/inducer and suppressor-inducer functions.
Assuntos
Anticorpos Monoclonais , Antígenos de Superfície/imunologia , Linfócitos T Reguladores/imunologia , Linfócitos T/imunologia , Formação de Anticorpos , Antígenos de Diferenciação de Linfócitos T , Células Cultivadas , Humanos , Ativação Linfocitária/efeitos dos fármacos , Mitógenos de Phytolacca americana/farmacologia , Linfócitos T/classificação , Linfócitos T Auxiliares-Indutores/imunologiaRESUMO
This report demonstrates that human peripheral blood T lymphocytes, triggered by allogeneic cells or soluble antigens, elaborate helper factor(s) that promotes the in vitro differentiation of TNP altered-self reactive human CTL. Helper factor(s) alone is not sufficient for the generation of these killer cells, but requires the presence of TNP-derivatized autologous stimulators during sensitization. Additional experiments were performed with antisera to a human Ia-like antigen, p23, 30. These studies indicate that human Ia-like antigens play an important role in both the induction of helper factor(s) and in the functional activity of preformed helper factor(s) molecules.
Assuntos
Citotoxicidade Imunológica , Isoantígenos , Nitrobenzenos/farmacologia , Linfócitos T/imunologia , Trinitrobenzenos/farmacologia , Relação Dose-Resposta Imunológica , Humanos , Cinética , Complexo Principal de Histocompatibilidade , Vacina contra Caxumba/imunologia , Solubilidade , Toxoide Tetânico/imunologia , Tuberculina/imunologiaRESUMO
Utilizing a PFC assay to quantitate the polyclonal activation of human peripheral blood B lymphocytes, we have investigated the induction and functional activity of MLC-derived human helper factor(s). Our data demonstrate that highly purified responder T cells, but not B or null cells, are required for the elaboration of MLC helper factor(s) that trigger the in vitro differentiation of B lymphocytes into PFC. Helper factor can trigger B cell maturation in the absence of helper T cells, since complement- (C) mediated lysis of the small (less than 5%) fraction of T cells present in anti-F(ab)2 immunoabsorbent column purified B cell population eliminates the PWM induced, but not the helper factor-induced PFC response. Responder T cells required for helper factor production do not bear surface membrane Ia, since alpha p23,30 + C treatment of this population does not affect helper factor generation. In contrast, alpha p23,30 + C treatment of the allogeneic stimulator cell population eliminates helper factor production. Taken together, these results demonstrate that interaction between Ia-bearing stimulator cells and Ia- responder T cells is required for the production of MLC-derived helper factor. In additional experiments, we determined that alpha p23,30, in the absence of C, totally abrogates the PFC response triggered by MLC helper factors. This result suggests an important role for Ia antigens in the functional activity of preformed helper factor molecules.
Assuntos
Linfócitos B/citologia , Antígenos de Histocompatibilidade , Linfócitos T/imunologia , Anticorpos , Células Produtoras de Anticorpos/imunologia , Diferenciação Celular , Células Cultivadas , Técnica de Placa Hemolítica , Humanos , Teste de Cultura Mista de Linfócitos , Mitógenos de Phytolacca americana/farmacologia , Toxoide Tetânico/imunologiaRESUMO
Human peripheral blood E+ lymphocytes were alloactivated by E- cells and then purified by repeat E+ selection. As described by others, alloactivated cells (E+d6) but not unactivated E+ cells were capable of stimulating both allogeneic and autologous mixed lymphocyte cultures (MLC). To further characterize the subset of cells responsible for this function, we used a variety of monoclonal antibodies to human T cells and monocytes. Treatment of E+d6 cells with anti-Ia OKI1) plus complement (C) abrogated their stimulating capacity. In contrast, treatment with the anti-T cell antibodies OKT3, OKT11A, OKT4, OKT8 plus C or anti-macrophage antibody (OKM1) plus C failed to eliminate their MLC stimulatory capacintibodies to human T cells and monocytes. Treatment of E+d6 cells with anti-Ia OKI1) plus complement (C) abrogated their stimulating capacity. In contrast, treatment with the anti-T cell antibodies OKT3, OKT11A, OKT4, OKT8 plus C or anti-macrophage antibody (OKM1) plus C failed to eliminate their MLC stimulatory capacintibodies to human T cells and monocytes. Treatment of E+d6 cells with anti-Ia OKI1) plus complement (C) abrogated their stimulating capacity. In contrast, treatment with the anti-T cell antibodies OKT3, OKT11A, OKT4, OKT8 plus C or anti-macrophage antibody (OKM1) plus C failed to eliminate their MLC stimulatory capacity. Because OKT3 recognizes the majority of T cells and OKT11A recognizes virtually all E+ cells, we reasoned that a contaminating non-T cell containing the MLC-stimulating capacity may be present within the alloactivated E+ population. To further address this question, E+d6 cells were positively and negatively selected using a rosetting technique with anti-Ig-coated red cells. The positively selected OKT3, OKT11A, OKT4, or OKT8 E+d6 cells retained minimal ability to stimulate MLC, whereas the corresponding negatively selected populations were highly enriched in this function. Phenotypic analysis of the isolated populations failed to demonstrate greater than 1% surface membrane Ig(SmIg) positive cells. Taken together, these results suggest that the MLC stimulatory capacity of alloactivated E+ cells is contained within an Ia+, OKM1-, SmIg- non-T cell subset.
Assuntos
Eritrócitos/imunologia , Antígenos de Histocompatibilidade Classe II/imunologia , Linfócitos/imunologia , Formação de Roseta , Anticorpos Monoclonais/imunologia , Antígenos de Superfície , Humanos , Teste de Cultura Mista de Linfócitos , Fenótipo , Receptores de Antígenos de Linfócitos B/imunologiaRESUMO
T-B and T-T interactions involved in the regulation of PWM-triggered human B cell differentiation were studied in vitro. Functionally distinct human T cell subsets were isolated by C-mediated lysis by using the monoclonal antibodies OKT4 and OKT8. Graded numbers of either untreated or irradiated T cell subsets were added to autologous B cells, and total antibody synthesis was measured after 5 to 6 days of culture by using a highly sensitive reverse hemolytic plaque assay. The data indicate that a) the helper activity that is exclusively contained within the OKT4+ population is radiosensitive. Only at high T/B ratios can this radiosensitivity be overcome; b) the OKT8+ population contains radiosensitive cells important in suppressing B cell differentiation, and c) the suppression induced with OKT8+ cells requires the presence of radiosensitive OKT4+ cells. Thus, OKT8+ cells added to cultures containing B cells and irradiated OKT4+ cells do not suppress the PFC response. Addition of unirradiated OKT4+ cells to these cultures permits reexpression of suppression by OKT8+ cells. It is concluded that two radiosensitive cells, one within the OKT4+ population and the other within the OKT8+ population, collaborate to induce suppression. Possible mechanisms for this suppressive interaction including induction of suppressor precursor cells within the OKT4+ population or inhibition of OKT4+ helper cells by OKT8+ cells are discussed.
Assuntos
Anticorpos , Linfócitos B/citologia , Cooperação Linfocítica , Linfócitos T/classificação , Antígenos de Superfície/imunologia , Linfócitos B/imunologia , Diferenciação Celular , Células Clonais/imunologia , Humanos , Linfócitos T/imunologia , Linfócitos T/efeitos da radiaçãoRESUMO
The relationship between immunoregulatory T-cell function and the expression of T-cell subset-specific differentiation antigens was examined using a phenotypically anomalous human T-cell line (TCL), termed H-1. H-1 cells were found to express T11, extremely high levels of T3, but no T4 nor T8 antigen. Despite their lack of T4 antigen expression, H-1 cells could be activated by coculture with pokeweed mitogen (PWM), anti-T3 antibody, or autologous B cells to provide potent help for B-cell differentiation into plaque-forming cells (PFC). In contrast, H-1 cells did not suppress the PFC response triggered by PWM-activated T4+ cells. These results demonstrate that the expression of the T-cell subclass-specific differentiation antigen, T4, is not required for a T cell to become activated and to implement the program for helper function. In addition, enhanced expression of T3 on the T4-, T8-, H-1 cell surface may reflect a compensatory upregulation of the T3/Ti receptor complex on T cells which are deficient in these nonpolymorphic associative recognition structures.
Assuntos
Antígenos de Superfície/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Antígenos de Diferenciação de Linfócitos T , Linfócitos B/imunologia , Diferenciação Celular , Linhagem Celular , Humanos , Ativação Linfocitária/efeitos dos fármacos , Cooperação Linfocítica , Fenótipo , Mitógenos de Phytolacca americana/farmacologiaRESUMO
In previous reports we have demonstrated that human T cells, responding to soluble and alloantigens, release helper factor(s) that amplify primary in vitro hapten-altered-self-reactive CTL responses. In the present studies, we have employed complement-fixing monoclonal antibodies (OKT4 and OKT8) that recognize functionally distinct human T cell subsets to investigate the role of T-T interaction in the generation of these killer cells. In all experiments, purified OKT4+ responder T cells were deficient in cytotoxic activity, whereas responder populations containing OKT8+ T cells generated substantial cytotoxicity; demonstrating that TNP-altered-self-reactive CTL precursors are contained within the OKT8+ T cell subset. Further, optimal cytotoxic responses were obtained from responder populations containing both OKT4+ and OKT8+ T cells, suggesting that cooperative interaction between these subsets may result in an amplification of killer cell activity. This interpretation was supported by the following observations: (1) the amplifying effect of soluble antigen required the presence of both OKT4+ and OKT8+ responders; (2) during MLC, OKT4+ but not OKT8+ responder T cells generate helper factor(s) that amplify TNP-altered-self-reactive CTL responses; (3) helper factor(s) bypass the requirement for direct OKT4-OKT8 T cell interaction, triggering a CTL response that is proportional to the percentage of OKT8+ T cells present within the responder population. In additional studies, we determined that the TNP-altered-self-reactive effector CTL maintain the OKT3+, OKT4-, OKT8+ surface phenotype displayed by the CTL precursor.