Long-term maintenance in vitro of human T cells by repeated exposure to the same stimulator cells. Differences when using repeated stimulation in allogeneic mixed leukocyte culture and when using stimulation with autologous lymphoblastoid cells.

Cells from one-way human mixed leukocyte cultures (MLC) which had reverted to small lymphocytes after 2 weeks' incubation responded with accelerated kinetics and higher thymidine incorporation on restimulation with lymphocytes or lymphoblastoid cell line (LCL) cells having relevant antigens. In contrast to fresh lymphocytes, they did not respond to autologous LCL cells. Cultures could be restimulated every second week with relevant allogeneic lymphocytes and could thus be maintained for periods of up to 4 months. Almost all these cultured cells had T-cell characteristics, during stimulation as well as in their reverted phase. The response to phytohemagglutinin (PHA) successively disappeared with repeated allogeneic restimulation, whereas the response to the relevant lymphocytes and cells of related donors was maintained. When lymphocytes had been stimulated with autologous LCL cells, the restimulation response was accelerated, although lower than after the primary stimulation. Restimulated cultures could not be maintained by further restimulation. Allogeneic and autologous LCL were equally efficient restumulators. A low level of stimulation was also achieved with allogeneic lymphocytes. The PHA response was usually reduced.     

Long-Term Maintenance In Vitro of Human T Cells by Repeated Exposure to the Same Stimulator Cells

Cells from one-way human mixed leukocyte cultures (MLC) which had reverted to small lymphocytes after 2 weeks' incubation responded with accelerated kinetics and higher thymidine incorporation on restimulation with lymphocytes or lymphoblastoid cell line (LCI.) cells having relevant antigens. In contrast to fresh lymphocytes, they did not respond to autologous LCL cells. Cultures could be restimulated every second week with relevant allogeneic lymphocytes and could thus be maintained for periods of up to 4 months. Almost all these cultured cells had T-cell characteristics, during stimulation as well as in their reverted phase. The response to phytohemagglutinin (PHA) successively disappeared with repeated allogeneic restimulation. whereas the response to the relevant lymphocytes and cells of related donors was maintained. When lymphocytes had been, stimulated with autologous LCL cells, the restimulation response was accelerated, although lower than after the primary stimulation Restimulated cultures could not be maintained by further restimulation. Allogeneic and autologous LCL were equally efficient restimulators. A low level of stimulation was also achieved with allogeneic lymphocytes. The PHA response was usually reduced.     

Some requirements for a linear cell dose response in vitro assay for the T-cell progenitors of cytotoxic lymphocytes.

To develop a precise assay for the T-cells progenitors of cytotoxic lymphocytes (CL-progenitors), lymphoid were cultured under optimal conditions in Marbrook vessels with mitomycin-treated allogeneic stimulator cells, and the total level of CL produced 5 days later estimated by a modified 51Cr release assay. Conditions were adjusted so an arithmetically linear cell dose response relationship was obtained. Three aspects of the cell dose response curve required attention. (1) At low responding cell inputs a macrophage-like cell became limiting (despite the presence of allogeneic macrophages in the stimulating cell population), leading to a lag in the response. This limitation was overcome by adding a low level of irradiated syngeneic macrophages, or by using irradiated syngeneic spleen 'filler' cells. (2) The slope of the resultant linear dose response region could be reduced if desired by changing from cellophane dialysis membranes to 0.1 mu pore size nuclepore membranes, suggesting a stimulatory role for some higher molecular weight soluble factor produced in the cultures. (3) At higher responding cell inputs a marked and extensive plateau was obtained. CL developing early in the response appeared to be destroying the allogeneic stimulator cells causing the response to be self-limiting. This problem was overcome by using a responding cell concentration lower than commonly employed. Assays using mixed leukocyte cultures in the lag or plateau regions could give misleading vlaues for CL-progenitor activity. It is suggested that some examples of apparent synergism in CL generation may have resulted from these effects, rather than T-cell helper T-cell progenitor interactions.     

Histocompatibility requirements for T cell help in specific in vitro antibody responses to influenza virus by human blood lymphocytes

Specific antibody responses to influenza virus were obtained in vitro from human blood mononuclear cells (PBM). The response was T cell-dependent, as shown by separation of PBM into E rosette-positive (E⁺) and -negative (E^?) populations. The histocompatibility requirements for T-B cell interactions in this response were analyzed by recombining E^? and E⁺ fractions from donors with varying degrees of HLA compatibility. No antibody formation was obtained from any allogeneic combination except for the special case of HLA identical siblings. As these experiments included combinations with shared or identical HLA-DR specificities, it was unlikely that genetic restriction alone could account for the failure of T-B cell collaboration. Evidence that suppression was responsible for the lack of antibody formation was obtained from experiments in which allogeneic E⁺ cells profoundly depressed specific antibody responses of intact PBM. In contrast, no such suppression was seen in pokeweed mitogen-driven polyclonal Ig synthesis for which there are no major histocompatibility complex requirements for T cell help. The suppressor activity of allogeneic E⁺ cells was found to be radiation-sensitive. By irradiating E⁺ cells, it was, therefore, possible to test for T cell help across an HLA barrier without unwanted suppressor effects. Under these conditions, (irradiated) E⁺ cells were able to collaborate with allogeneic E^? cells even with no HLA alleles in common. This was true even when autologous monocytes were depleted from the helper E⁺ population. Supernatants collected from antigen-driven cultures of allogeneic E^? and E⁺ cells were able to replace helper T cells in the specific antibody response to influenza virus. The apparent lack of genetic restriction in these responses might, therefore, be explained by the production of a nonrestricted helper factor.     

Antigen specificity and frequency of autologous and allogeneic helper T cells in the in vitro production of antibody against influenza virus by human blood lymphocytes

The frequency of antigen-specific helper T cells in human peripheral blood mononuclear cells was determined by limiting dilution analysis of specific in vitro antibody responses to influenza virus A/X-31 (A-H3N2). Limiting numbers of irradiated E rosette-forming (E⁺) (T) cells were added to a constant number of syngeneic non-rosette-forming (E^?) (“B”) cells and their ability to support production of antibody to influenza virus determined. The frequency of T helper cells was then calculated by a Poisson distribution analysis and found to range between 0.78 × 10^?5 and 2.86 × 10^?5. Specific antibody production can be obtained in this system from allogeneic combination of E^? and E⁺ cells provided that the added E⁺ cells are irradiated to abrogate alloactivated T suppressor effects. Limiting dilution analysis applied to determine the frequency of helper T cells under these conditions showed that in most, but not all, cases the frequency of T helper cells was higher in allogeneic combinations. This result could be interpreted as showing an increase in the number of activated specific T helper cells, due perhaps to a positive allogeneic effect. On the other hand, it could also be explained by the alloactivation of an entirely different subset of nonspecific helper cells, or by the production of a nonspecific allogeneic helper factor. The specificity of T cell help in allogeneic combinations was therefore examined by adding limiting numbers of E^? cells to a fixed number of E^? cells and simultaneously challenging with two non-cross-reacting influenza viruses A/X-31 and B/HK. Under these conditions, individual cultures produced antibody to A/X-31, B/HK or both. The frequency of cultures producing antibody to both viruses was as predicted for the chance occurrence of specific helper T cells for both antigens being present in the same culture. The fact that a significant number of individual cultures made antibody to only one antigen in both autologous and allogeneic combinations showed that T cell help was antigen-specific in both situations. Thus, for human in vitro antibody responses, antigen-specific T cell help can be obtained across a major histocompatibility (complex) barrier.     

Antibody formation by human tonsil cells in vitro.

The ability of human tonsil lymphocytes to give an anti-SRBC response in conventional cultures and in microcultures was studied. It was found that about two-thirds of tonsils responded with a significant number of plaque-forming cells (PFC), and that in some instances the response could be augmented if allogeneic tonsil cell (irradiated or intact) were added. Moreover, tonsil lymphocytes which failed to give a response on their own often responded upon addition of an appropriate number of allogeneic tonsil cells. The response was remarkably improved if allogeneic tonsil supernatant or conditioned medium were added. An anti-SRBC response was also obtained if SRBC was omitted from the cultures. The frequency of anti-SRBC specific B cells was estimated as 1/60000 (f = 1-7 X 10(-5)).     

Antigen-specific suppression of human antibody responses by allogeneic T cells. II. Cell interactions involved in the generation of suppression

Specific antibody responses to influenza virus were obtained in vitro from human blood mononuclear cells (PBMC). Antibody production in these cultures was profoundly suppressed by the addition of allogeneic T cells with the surface phenotype Leu2a+ (CD8+), Leu8-. Suppression by allogeneic T suppressor (Ts) cells required interactions only between T-depleted B (E-) cells and allogeneic Leu2a+. No evidence was obtained for T-T cell interactions, or for Ts inducer cells similar to those described for nonspecific antibody responses to pokeweed mitogen. Moreover, allogeneic E+, or allogeneic Leu2a+ cells were able to suppress specific antibody responses by E- cells when help was provided by T cell-replacing factor showing that the target of suppression was the responding E- cells, and not T helper cells. In contrast to allogeneic T cells, allogeneic E- cells did not suppress antibody production when added to cultures of unfractionated PBMC (E- + E+). That is, Ts cells activated to allogeneic E- were unable to suppress antibody production by the syngeneic E- cells present in the same culture tube. This result shows that alloactivated Ts cells were specific for the allogeneic E- target cells, and that suppression was not mediated by nonspecific allogeneic effects. Allogeneic Ts cells therefore differ from Ts cells in pokeweed mitogen responses by their specificity, and by their activation in the absence of Ts inducer cells.     

Regulation of the primary in vitro antibody response in human peripheral blood lymphocytes: different effects of mitogen-induced and spontaneous T suppressor cells.

The specific response of human peripheral blood lymphocyte cultures to TNP-polyacrylamide was suppressed by the addition of concanavalin A (Con A). A dose of Con A (0.5 microgram/ml) could be selected, which induced a reproducible but incomplete suppression independent of the magnitude of the anti-TNP response. Con A-stimulated cells could transfer the suppression to autologous or allogeneic responding cells. The suppressor activity was present in the E-rosette forming cell fraction and was abolished by mitomycin C treatment prior to incubation. With a particular batch of foetal bovine serum, spontaneous suppressor cells were produced which suppressed the response of autologous and allogeneic lymphocytes. In allogenic mixtures, the otherwise enhancing allogeneic effect was replaced by a marked suppression from spontaneous suppressor cells. This suppression was higher than that exerted on autologous lymphocytes, suggesting that an unexpected negative allogenic effect had taken place. Spontaneous suppressors were ineffective when added on day 2 of a culture responding to TNP-polyacrylamide, whereas Con A induced suppressors were fully effective.     

Immunoregulatory Function of Human Cord Blood Lymphocytes on Immunoglobulin Production

ABSTRACT: The functional maturity of human umbilical cord blood B lymphocytes and the immunoregulatory activity of cord T lymphocytes were assessed by measuring the in vitro immunoglobulin production by B cells from either cord or adult blood. Supernatants from 48-hr pokeweed-mitogen (PWM) stimulated cord or adult lymphocyte cultures were added to cord or adult B cell cultures in the presence of PWM; a significant amount of immunoglobulin was produced in adult B cell cultures only. Adult B or T cells were then cocultured with cord T or B cells; a significant amount of immunoglobulin was again found only in adult B cell cultures. These results indicated that cord B cells were functionally immature and that cord helper T cell function was adequate but masked by excessive suppressor activity. Indeed, addition of cord T cells but not of allogeneic adult T cells to PWM stimulated adult lymphocyte cultures inhibited their immunoglobulin production; this confirmed cord T cells' increased suppressor activity. Cord T cells were not intrinsically suppressive since they failed to suppress immunoglobulin production by Epstein-Barr Virus (EBV) transformed B cells. They could be activated, however, by PWM or allogeneic cells (in mixed lymphocyte cultures) and their effect was mediated via soluble factor(s) as demonstrated by the suppressor effect of these culture supernatants on immunoglobulin production by unfractionated adult lymphocytes. In contrast, when these supernatants were added to T cell-depleted adult lymphyocyte cultures, enhancement rather than suppression was observed. These results indicated that the soluble factor(s) released by Cord T lymphocytes was not suppressing per se but induced suppression through activation of suppressor cells.     

Specific allogeneic help by T lymphocytes from patients with systemic lupus erythematosus.

Unfractionated mononuclear cells from patients with systemic lupus erythematosus (SLE) immunized with influenza vaccines do not produce a secondary in vitro anti-influenza antibody response when challenged with virus antigen. Irradiated T lymphocytes from normal, disease control and from SLE donors whether vaccinated or not, help allogeneic normal non-T cells to produce specific anti-influenza antibody in vitro. Irradiated normal T cells, however, do not help allogeneic non-T cells from SLE donors. Non-irradiated T cells from 40% of the SLE patients, irrespective of whether or not they had been vaccinated, also provide specific help for MLC incompatible normal non-T cells in the influenza antibody response. This non-restricted interaction was not seen using non-irradiated T cells from any normal or disease control donor. No anti-DNA antibodies were produced in virus stimulated cultures of non-irradiated or irradiated SLE T cells with allogeneic normal non-T cells.     

Suppression of the generation of secondary virus-specific proliferative and cytotoxic T lymphocytes by suppressor cells induced during primary anti-viral sensitization in vitro

Murine spleen cells first primed with syngeneic vaccinia virus-infected peritoneal exudate cells (PEC) in vitro and then restimulated with the virus failed to give a typical virus-specific secondary cytotoxic T lymphocyte (CTL) response. In contrast, "memory' spleen cells from mice primed with the virus in vivo produced CTL after the same challenge with virus-infected PEC in vitro. In the former situation, the lack of a virus-specific secondary CTL response by in vitro primed and restimulated spleen cells seemed to be associated with the generation of suppressor cells in cultures; these cells inhibited the cytotoxic as well as proliferative secondary and tertiary responses of spleen presensitized with virus in vitro alone, or in vivo plus in vitro. Weak suppressor activity was also induced in control spleen-cell cultures from normal unprimed or virus-primed mice that were not stimulated with virus-infected cells, suggesting either a quantitative difference in the generation of suppression or, alternatively, the co-existence of virus-dependent and independent suppressor cells in the virus-stimulated cultures. Our experiments cannot conclusively establish that suppression is T-cell mediated and/or possibly natural-killer-(NK)-cell dependent. The suppressor phenomena were exerted by irradiation resistant (850 rad) lymphocytes that passed through nylon wool columns and were sensitive to treatment with anti-Thy-1 antibody plus C; but the suppressor cells were partially reactive across allogeneic barriers.     

The regulation of T cell responses by spontaneously active suppressor cells.

In studying T cell regulation, peripheral blood mononuclear cells from normal subjects were examined for 'spontaneous', rather than mitogen-induced, suppressor cell activity. Normal blood leucocytes from 30 subjects included a subpopulation of cells capable of suppressing the response of lymphocytes to the T cell mitogen phytohaemagglutinin by 21-35%. The indicator system for these studies consisted of fresh normal lymphocytes stimulated by three concentrations of PHA in the presence or absence of normal but mitomycin C treated peripheral blood lymphocytes. To measure accurately the spontaneous suppressor cell activity, additional cultures were needed to control for the suppressive effects of crowding and metabolic competition. Allogeneic, cryopreserved 'B' cell enriched populations, supplied satisfactory control cells for this purpose. While allogeneic culture systems could induce significant suppressor cell activity after 7 days of co-culture, they could not induce this activity in the 3 days required to assay spontaneous suppressor cell effects. In developing this assay we noted that (a) crowding became a factor in the cellular response to mitogens with concentrations higher than 2 X 10(4) cells/well, (b) spontaneous suppressor cell activity decreased rapidly once cells were placed in culture and (c) both spontaneous and concanavalin A (Con A) activated suppressor cells could significantly reduce the response to PHA even when added to cultures established with mitogens 72 hr earlier. The ability to measure spontaneous suppressor cell activity in vitro will allow more physiological studies of the membrane markers and functional characteristics of these cells than is possible in conventional studies utilizing Con A. In addition, this assay allows the detection of enhanced in vivo activity of suppressor cells not easily detected in assays relying on mitogen induction of suppression. Such increased activity is thought to be an important factor in the pathogenesis of a number of human diseases.     

Specific antibody responses by high- and low-density human peripheral blood B cells: T-helper cells and T-cell replacing factor (TRF) act on different B-cell subpopulations.

Antibody production to influenza A strain virus X31 (H3N2) was measured in cultures of peripheral blood mononuclear cells (PBMC) stimulated with either antigen (X31) or pokeweed mitogen (PWM). With some donors, X31 antibody was produced in response to antigenic stimulation, but not as part of the polyclonal response to PWM, suggesting that antigen and PWM may be acting on different B-cell subpopulations. To test this hypothesis, T-cell depleted PBMC (E-) cells were fractionated on discontinuous Percoll gradients and assayed for antibody production in response to antigen or PWM. Fraction I (FrI = SG less than 1.070) cultured in the presence of T cells responded well to PWM, but not at all to X31. FrII (1.070 less than SG less than 1.075) and FrIII (SG greater than 1.075) cultured in the presence of T cells both responded well to X31, but only the medium-density B cells (FrII) were able to make specific antibody when T cells were replaced with T-cell replacing factor (TRF). Specific X31 antibody responses by medium- and high-density B cells (FrII and FrIII) were suppressed equally by the addition of allogeneic T-suppressor (Ts) cells. When allo-activated Ts cells were inactivated by irradiation, allogeneic T-helper (Th) cells were able to collaborate with both FrII and FrIII B cells in specific antibody responses to X31. Since TRF was not able to substitute for T cells in specific antibody responses by FrIII B cells, this result shows that allogeneic T-cell help was not mediated by non-specific 'allogeneic effect' factors and apparently requires cognate T cell-B cell interactions.     

Continuously proliferating allospecific T cells, lifespan and antigen receptors.

Allospecific mouse T cells derived from one-way mixed lymphocyte cultures after 19 months in tissue culture still require allogeneic stimulation for their proliferation. If injected into normal or immunodeficient mice these cells do not form tumors. After proliferation over long time periods or after extensive expansion these T cells tend to lose their cytotoxic activity which cannot be reconstituted by addition of cells from active cultures of a different specificity. Accordingly, the specificity of the cytotoxic and proliferative response was investigated. The cytotoxic activity appears to be specific for the H-2 SD antigens, while the proliferative response and also the activation of cytotoxicity in resting cultures is only triggered by lymphocyte activating determinants (LAD) coded either inside or outside the H-2 region. The receptors for LAD on these allospecific T cells appear to be clonally distributed. This finding comes from the observation that old and therefore highly selected lines only react to some LAD while younger ones react to many others as well. A search for immunoglobulin on these functional and antigen-specific thymus-derived lymphocytes (T cells) was performed using surface radioiodination, immunoprecipitation and polyacrylamide gel electrophoresis combined with autoradiography. Using various antisera no or very little immunoglobulin molecules were found on these T cells. The small amount of immunoglobulin was attributed to a contamination of allogeneic spleen cells used to stimulate the T killer cells.     

Function,target-cell preference and cell-surface characteristics of herpes-simplex virus type-2-induced non-antigen-specific killer cells

Wild-type and congenitally athymic nude mice injected with herpes-simplex virus type 2 (HSV 2) responded with a local outburst of non-antigen-specific killer cells masking any virus-specific response. Cytolytic activity could be assayed on mouse-tumor cell lines and on syngeneic or allogeneic non-transformed cells from various sources. Some of the tumor cell lines and proteose-peptone-induced peritoneal exudate cells were lysed more efficiently after infection with either HSV 2, vaccinia or influenza A virus. Preference for virus-infected target cells was already expressed 24 hours after HSV -2 injection. Killing activity was not H-2- restricted, not complement- or immunoglobulin-dependent and did not involve Fc receptors. The cytotoxic cells were non-adherent and could be shown to express Thy1, Quat4, and Quat5 cell-surface antigens. They lacked immunoglobulin and Lyt1 : Lyt2,3 determinants. The functional and serological characteristics identify the HSV-2-induced cytolytic cells as natural killer (NK) cells. The potential importance of this cell population for natural resistance will be discussed.     

Human B cell function in responder and non-responder individuals. II. The role of T helper cells in promoting the PWM-induced B cell production of immunoprotein

Sorted OKT4+ cells treated with pokeweed mitogen (PWM) and subsequently X-irradiated were used as a source of helper T cells to examine human T and B cell function. PWM-induced immunoprotein synthesis by human peripheral blood lymphocytes was the model used to study the cellular interactions. PWM was shown to induce helper T cell function which caused non-PWM treated B cells to secrete immunoglobulin. PBL from certain individuals could not be induced by PWM to secrete Ig therefore allogeneic co-cultures of helper T cells and B cells were examined to define the defective cell population. Ig synthesis in allogeneic cultures of T and B cells was always greater than that observed in autologous cultures when cells from responders were assayed. However, when allogeneic cultures were initiated using B cells from a responder and PWM treated T cells from a non-responder and examined for Ig synthesis, the B cell responses were markedly lower than seen in the autologous responder cultures. In addition, PWM activated helper T cells from a responder induced a significantly higher Ig synthesis by B cells from a non-responder. These observations indicate that PBL from individuals who do not respond in a PWM driven Ig synthesis assay have relatively normal B cell function but are deficient in helper T cell function.     

Age-related impairment of the in vitro antibody response in the human.

We have used trinitrophenyl polyacrylamide beads (TNP-PAA) to induce a primary in vitro antibody response toward TNP in cultures of peripheral blood lymphocytes (PBL) from aged individuals. The response was virtually non-existent whereas PBL from young control individuals were able to respond. Recombination experiments showed that: (1) aged PBL did not suppress the response of young PBL; (2) young T cells enhanced the response of aged (unfractionated or T-depleted) cells; (3) aged T cells were unable to restore the response of young T-depleted cells, in contrast to young allogeneic T cells. Upon stimulation with Concanavalin A (Con A) aged PBL displayed a moderately diminished proliferative response and a normal ability to suppress the anti-TNP response of autologous PBL (in the few responders). However, contrasting with young PBL they could not suppress the anti-TNP response of young allogeneic PBL.     

Production of Macrophage Migration Inhibition Factors by Virus-Infected Cell Cultures

Macrophage migration inhibitory (MIF-like) activity was demonstrated in the supernatant fluids from cultures of African green monkey kidney cells (BGM) infected with mumps virus or Newcastle disease virus. We could detect no such activity in noninfected cultures. The virus-induced activity reported here is not due to nonspecific cytotoxic material released by dead or dying cells, and it does not require cell replication for its production. Preliminary estimates of molecular weight by Sephadex G-100 chromatography revealed a broad band of activity associated with the 45,000 and 65,000 markers. These are significantly smaller than previously reported chemotactic substances from virus-infected cultures, and thus appear to represent different cell products. These MIF-like factors may be produced concomitantly with interferon. However, ultraviolet irradiation of appropriate duration abolishes the ability of viruses to induce substances with MIF-like activity while preserving the ability to induce interferon. This strongly suggests that interferon is not the agent responsible for the macrophage migration inhibition effect. The functional properties of these various cell products induced by virus infection suggest that they all may play a role in the response to virus infection in vivo.     

Plaque-Forming Cells in Man

We have investigated the ability of allogeneic, irradiated T lymphocytes to induce proliferation and immunoglobulin (Ig) secretion in untreated peripheral blood B lymphocytes. Non-mitogen-activated co-cultures of isolated T and B lymphocytes from selected, full-house HLA-A,B and D/DR antigen-phenotyped donors were reconstituted in a ratio of 4:1. Proliferation was assessed on day 5-6 of culture by the 3H-thymidine incorporation technique, and the Ig secretion was monitored on day 6 with a protein A plaque-forming cell (PFC) assay. B lymphocytes were able to differentiate into PFC, and the number of plaques was significantly higher in cultures of cells with two HLA-D/DR antigen incompatibilities than in those sharing one antigen. In cultures of peripheral blood lymphocytes with no HLA-D/DR antigen difference, only a few PFC developed. HLA-A and B antigens had no influence on the response. Further, monocytes were not an absolute requirement for allogeneic activation of B cells. Sonicated T cells and culture supernatants from allogeneic T- and B-cells cultures were not able to induce PFC formation in B lymphocytes. Our results indicate that the PFC response obtained in non-mitogen-activated cultures of allogeneic T and B lymphocytes is dependent on HLA-D/DR disparity or on genes encoded in the HLA-D/DR region.     

Autoreactive cytotoxicity in HIV-infected individuals

A possible role for autoimmunity in the pathogenesis of HIV infection has been suggested, based upon the certain degree of homology shared by HIV gp41 and MHC class II molecules. A number of humoral markers of autoimmunity have since been found in seropositive subjects. We have evaluated the cellular autoreactive response in HIV-infected individuals. Our study demonstrates the existence of a cytolytic activity, present in seropositive but not in seronegative subjects. This activity is mediated by CD3+ T cells, which only occasionally express the CD8 or the CD4 surface markers. Effector cells do not appear to exert their activity in a MHC-restricted fashion, since allogeneic target cells could also be killed, recovered from allogeneic seropositive as well as from seronegative subjects. Several types of target cells were lysed: T cell blasts and Epstein-Barr virus (EBV) transformed B cells, suggesting that the target antigen is common to at least these two cell types. The fact that cells from seronegative individuals were lysed argues against the recognition of an HIV-specific antigen. The nature of the target determinants and the identity of the effector cells are discussed.