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Larvae of the Northern pine processionary moth (Thaumetopoea pinivora, TP) carry microscopic needles (setae), which by penetrating skin and mucous membranes, may cause inflammatory/immune derived symptoms in man. In the present study the stimulatory effects of setae on human blood lymphocytes in vitro was investigated. Blood mononuclear cells were separated from venous blood or buffy coat of ten healthy individuals, six previously exposed to setae and four with no known exposure. Lymphoproliferation was measured as uptake of 3H-thymidine. Setae were prepared from TP larvae. Setae and saline setae extracts stimulated proliferation of T-lymphocytes in the presence of monocytic cells. Stimulation was pronounced in cells from persons who had been exposed to setae, and weak in cells from non-exposed donors. Chitin also induced lymphocyte proliferation in most donors, but to a lesser extent and independently of donor's previous exposure to setae. In conclusion, setae contain molecules that in the presence of monocytes activate human T-lymphocytes to proliferation. The antigenic nature of stimulatory molecules was supported by the significantly stronger lymphocyte response in persons previously exposed to setae than in non-exposed donors. The nature of such molecules remains to be defined.

We investigated whether the described immune evasion of Epstein-Barr virus (EBV)-infected malignant Hodgkin and Reed-Sternberg (HRS) cells in Hodgkin's disease (HD) is paralleled by a disturbed expression of the signal transduction molecule zeta associated with CD3 and CD16 in tumour-associated T lymphocytes (TAL). Flow cytometric analysis revealed a significantly lower zeta expression in CD3+/4+, CD3+/8+ and CD16+ patient peripheral blood lymphocytes (PBL; n = 10) compared with normal donor PBLs (n = 11). When patient PBLs were compared with the corresponding TAL, the latter showed a significantly higher (CD3+/4+) or equal (CD3+/8+) zeta expression. The EBV status of the tumours did not correlate with zeta expression in the TAL. Immunohistochemical staining revealed zeta-positive lymphocytes among the adjacent bystander cells of the HRS cells in all analysed tumours (n = 8), irrespective of tumour EBV status. In conclusion, these results do not support downregulation of zeta in TAL as a critical mechanism contributing specifically to the immune escape of EBV+ HRS cells.

Epstein-Barr virus (EBV) expression was investigated by immunohistochemistry (latent membrane protein 1 [LMP-1]) and in situ hybridization (EBV encoded RNA [EBER]) in biopsies from 95 patients with untreated Hodgkin's disease (HD). Tumour EBV status was related to EBV antibody titres, spontaneous and concanavalin A induced blood lymphocyte DNA synthesis, serum levels of soluble (s) CD4, sCD8, sCD25, sCD30, sCD54, beta2-microglobulin, thymidine-kinase, routine chemistry, patient characteristics, complete remission and survival. The median follow-up time was 145 months (range 60-257). Tumour EBV-positive (n = 30; 33%) and negative (n = 62; 67%) patients did not differ with regard to sex, age, stage, presence of bulky disease or B-symptoms, remission rate or survival. The proportion of EBV+ cases was significantly higher among patients with mixed cellularity histopathology (58%) as compared to the nodular sclerosis subtype (18%; P < 0.001). The total white blood cell (WBC) counts were significantly lower in EBV+ patients (P < 0.01), who also had significantly higher levels of sCD54 (P < 0.02) and a tendency towards lower levels of sCD30 (P = 0.056). Patients in the tumour EBV+ group had significantly higher IgG antibody titres to restricted early antigen (EA-R) (P < 0.02). Hence, clinical features and outcome were not related to tumour EBV status. However, HD patients with EBV+ tumours had elevated sCD54 levels, higher antibody titres to EA-R and decreased total WBC counts. A potential causal relationship between EBV tumour status and these findings needs to be further explored.

Measurement of the soluble form of CD8 antigen (sCD8), a surface membrane component of suppressor/cytotoxic T cells, has yielded useful information relevant to prognosis in children with acute lymphoblastic leukaemia and Hodgkin's disease (HD). An ELISA technique was used to measure the amount of sCD8 in sera from 123 adults with untreated HD. Significantly higher mean sCD8 levels were found in patients with advanced disease (stage III-IV; P less than 0.001), B-symptoms (P less than 0.001), male sex (P less than 0.05) and increased spontaneous and decreased Concanavalin A induced blood lymphocyte DNA-synthesis (P less than 0.05). Actuarial survival of patients with high sCD8 levels was significantly worse than that of the remainder (P less than 0.05). However, the sCD8 level did not add prognostic information to that achieved by age, sex, lactate dehydrogenase (LDH) or clinical stage. A significant correlation between the sCD8 and LDH levels (r = 0.33; P less than 0.001) and inverse correlations between sCD8 levels and total blood CD4+ (r = -0.52; P less than 0.001) and CD3+ (r = -0.39; P less than 0.01) cell counts were found. Ten patients were also studied in complete remission, showing a significantly reduced sCD8 level in comparison to the pretreatment value (P less than 0.05). Increased sCD8 in HD may indicate enhanced suppressor T cell activity possibly compromising the host disease balance which could explain the association with prognosis.

The study comprises 26 patients with non-Hodgkin's lymphoma who entered complete remission. Lymphocyte subsets and spontaneous and mitogen (ConA, PWM)-induced DNA synthesis were recorded in purified blood lymphocytes harvested before treatment, in remission and in relapse. The T and B lymphocytopenia noted in untreated patients persisted in complete remission. At diagnosis the spontaneous DNA synthesis was increased and the response to mitogen was decreased as compared to controls. Both functions became normalised during complete remission in most patients under 60 yr. The lymphocyte abnormalities reappeared during relapse. A low response to mitogen (ConA) in complete remission seemed to predict early relapse in patients younger than 60 yr. It is concluded that functional abnormalities of blood lymphocytes in non-Hodgkin's lymphoma are closely related to the presence of active disease.

We investigated the possibility of using a standard electronic laboratory cell counter, of a type which permits manual settings of discriminator and sensitivity levels, for the quantitation of E rosette forming human blood lymphocytes. The electronic counting was performed on rosette specimens prepared from 10 healthy donors, nine lymphoma patients and eight patients with chronic lymphocytic leukaemia (CLL) of B cell type. The simple procedure which was earlier developed for the electronic counting of thymocyte rosettes in guinea-pigs was used and now supplemented with a method for rapid, graphical determination of rosette frequency. The low level of E rosettes in CLL patients was easily distinguished from the higher level seen in the other groups and there was a good agreement between electronic and visual determinations. In samples with a high rosette frequency, the cell counter initially gave 10-30% lower values than when recorded visually. This discrepancy was corrected, however, by a minor adjustment of discriminator level and the introduction of a correction procedure for the presence of double rosettes. We conclude that the electronic counter can well be used for the counting of human rosette forming blood lymphocytes and, due to a high precision and large capacity, this approach would prove to be useful when screening of a large number of samples is required.

We studied the immunocompetence of 6 healthy twins, whose monozygotic or dizygotic same-sexed twin partner had died from Hodgkin's disease. Lymphocyte DNA synthesis induced by concanavalin A was markedly reduced at 3 different concentrations in all twins compared to an age-matched group of healthy controls. The lymphocyte response to pokeweed mitogen and to phytohaemagglutinin was also impaired. PPD induced lymphocyte DNA synthesis was low in 3 twins and correlated well with their delayed skin hypersensitivity to the antigen. One twin was completely anergic to 3 different skin antigens. The mean total blood lymphocyte count did not differ from that of controls. There was no change in T or B-lymphocyte subpopulations. The presence of a functional lymphocyte deficiency in all twins strongly suggests that the immunodeficiency in Hodgkin's disease is partly caused by genetic and/or environmental factors.

The effects of delayed cutaneous hypersensitivity (DCH) testing (Multitest, Institute Mérieux) on lymphocyte counts and functions were studied in seven healthy women. All had normal DCH responses and lymphocyte functions. No influence of the DCH testing on subsequent tests for lymphocyte subpopulations and mitogen- and antigen-induced DNA synthesis was observed. The PPD skin test did not boost or otherwise alter the in vitro lymphocyte response to PPD. It may be concluded that Multitest is a simple way of assessing DCH and that this test does not interfere with repeated in vitro tests of lymphocyte capacity.

Blood lymphocytes from thirty-nine untreated patients with Hodgkin's disease were tested for stimulatory and responding capacity in mixed lymphocyte culture. The response to allogeneic lymphocytes was impaired in ten patients but the mean response of the patient group did not differ from healthy controls. Lymphocytes from female patients responded better than male patients (P less than 0-02). The stimulatory efficacy was severely impaired in patients' lymphocytes (P less than 0-001). Age, clinical stage and histopathology had no significant influence on the responding or stimulatory capacity.

Blood lymphocyte functional capacity and serum immunoglobulins were studied in 40 patients with Hodgkin's disease (HD) admitted to Radiumhemmet, Stockholm, before treatment and in complete remission 2-56 months following termination of radiotherapy (total nodal irradiation [TNI]; n = 29) or chemotherapy (MOPP; n = 11). Lymphocyte studies included determination of total lymphocyte and T-cell counts and evaluation of spontaneous DNA synthesis during the first day of culture and mitogen-(concanavalin A, pokeweed mitogen) and antigen (purified protein derivative, PPD)-induced activation on the third day. Blood lymphocyte and T-cell counts decreased dramatically following TNI. A slow restitution was seen, but pretreatment levels were not reached even four years following therapy. The responses to ConA and PPD but not PWM were significantly reduced shortly after TNI. The mitogen response did not increase with time as did the PPD response. Lymphocyte counts and lymphocyte stimulation, which were severely depressed before treatment of patients in the chemotherapy group, remained unchanged 2-36 months after termination of therapy. A significant reduction of IgM levels was observed regardless of the mode of treatment. Splenectomy prevented the profound reduction of blood lymphocyte and T-cell counts following therapy but did not influence the other immunologic variables under study.

In a prospective study, sixty-five histologically verified Swedish cases of Hodgkin's disease were HL-A typed. The antigen frequencies in the patient group did not deviate from a group of normal controls. The frequency of HL-A8 was slightly increased in patients over 40 years (P less than 0.05). HL-A12 was markedly increased in patients below 40 years with a favourable histopathology (P less than 0.001). No association between sex or clinical stage and distribution of HL-A phenotypes was observed. The number of T-lymphocytes in blood and the spontaneous DNA synthesis and DNA synthesis induced by concanavalin A, pokeweed mitogen, phytohemagglutinin and PPD antigen in blood lymphocytes in vitro were determined in 60 patients. A significant increase of HL-A28 was noted in patients with advanced disease (P less than 0.01) and in patients responding poorly to PPD (P less than 0.001) and PWM (P less than 0.01). No correlation between HL-A antigen distribution and T-lymphocyte counts was observed.

Following indium-111 oxine-labelled autologous lymphocyte infusion, 39 lymphocyte scintigraphies were performed in 35 patients with Hodgkin's disease (HD). Lymphocytes were obtained after leukapheresis and Lymphoprep gradient centrifugation and were further purified by an adherence step to eliminate monocytes. A median of 1.2 (0.2-3.7) X 10(9) cells were labelled with 6.7 (range 1.9-16.6) Mbq indium-111 oxine and reinfused to the patients. Gamma camera imaging was performed with a GE 400 AT. An accumulation of radioactivity was seen at 54 of 61 sites with enlarged lymph nodes. Increased radioactivity was also seen at 16 sites with no previous clinical evidence of tumour involvement. These data seem very promising and with further methodological improvement lymphoscintigraphy may prove to be an important complement in the staging procedure and follow-up of patients with HD.

One hundred and twenty seven consecutive and previously untreated patients with Hodgkin's disease (HD) (mean age 47 years) from the Stockholm area admitted to Radiumhemmet, Karolinska Hospital, were studied. The age-matched control group consisted of 167 healthy adults. Incorporation of [14C]-dT was measured on Day 1 in unstimulated monocyte-depleted lymphocyte cultures, and on Day 3 in cultures activated by PWM, ConA and PPD, T and B cells were enumerated by surface markers. The patients had significantly decreased relative and total T-cell counts, and the lymphocyte DNA synthesis induced by mitogens and PPD was severely impaired, whilst the spontaneous DNA synthesis was significantly greater than in controls. At follow-up (mean 4 years) 40 patients have died. Deceased patients showed greater spontaneous lymphocyte activation and less response to mitogen and antigen stimulation than the survivors. The 5-year survival of patients with severe lymphocyte impairment was 20%, compared to 80% for the remainder. The lymphocyte tests added prognostic information to that from clinical staging. Disregarding the lack of knowledge of the mechanisms underlying the lymphocyte impairment, we suggest that these relatively simple immunological tests should be included in the clinical evaluation of HD patients and would guide the choice of therapy.

Possible candidates for hemic stem cells were studied in a quantitative investigation of the bone marrow of growing guinea pigs. The stem cell candidates were classified as lymphoid cells of types I-IV, roughly corresponding to small lymphocytes, large lymphocytes, micromyeloblasts and myeloblasts of other authors. The cellular content of the bone marrow of guinea pigs of three different ages was determined. All types of nucleated cells per femur were found to increase with age. When the cellular content was related to the body weight, a peak number of lymphoid cells was found in the intermediate age group with the exception of the lymphoid cells of type III, which reached its peak in the oldest animals. This cell type has been suggested by Dicke et al. as the probable stem cell responsible for colony formation.

The content of lymphocytes in blood samples from the splenic vein and the splenic artery of guinea pigs was determined and the veno-arterial difference in number of cells was used as measure of the net release of cells from the spleen into the blood. The splenic release of lymphocytes was reduced after whole body irradiation with 300 rad. This reduction could partly be prevented by transfusion of bone marrow cells after irradiation, 10(6) cells being the most effective dose. In thymectomized, irradiated animals the restoration of the splenic release of lymphocytes after irradiation was impaired in comparison with sham-operated animals. In the thymectomized animals the transfusion of 10(6) thymus cells had a restitutive effect on the splenic release of lymphocytes, while transfusion of bone marrow cells or daily treatments with thymosin, alone or combined with transfusion of bone marrow cells, had no such restitutive effect. The results indicate that restoration of the splenic release of lymphocytes into the blood after irradiation is thymus dependent and probably caused by a traffic of lymphocytes from the thymus to the spleen.

Lymphocyte counts and functional competence of lymphocytes from arterial and venous splenic blood were studied in six patients with Hodgkin's disease subjected to splenectomy. One patient was untreated, four were tested after mantle field treatment and a sixth patient had a splenic relapse after total nodal radiotherapy. The percentage of E binding cells in splenic venous blood was lower than that of arterial blood though no significant differences were found in total lymphocyte or E binding cell counts. The spontaneous lymphocyte DNA synthesis was lower in venous than in arterial splenic lymphocytes in all patients. Pokeweed mitogen (PWM)-induced DNA synthesis was much lower in lymphocytes from splenic venous blood than in those from arterial blood in two patients and marginally decreased in another two. The pattern of concanavalin A response was similar to that of PWM. The elimination of lymphocytes over the spleen could not be related to the presence of lymphocytotoxic serum factors or to splenic weight or histologically verified tumour involvement. The results support the notion that some facets of the blood lymphocyte abnormalities in Hodgkin's disease may be explained by removal of functionally active lymphocyte subpopulations in the spleen. It is also concluded that spontaneously activated lymphoid cells are detained in the spleen.

Purified blood lymphocytes from 81 consecutive and previously untreated patients with Hodgkin's disease were studied in vitro. The patients were evaluated 23 to 52 months after institution of therapy and were divided into two prognostic groups: 1. Complete remission with or without previous relapse/relapses. 2. Uncontrolled relapse or decreased from Hodgkin's disease. Lymphocyte activation by concanavalin A, pokeweed mitogen or phytohaemagglutinin was impaired and spontaneous DNA synthesis was high in patients with poor prognosis as compared to the good prognosis group. The prognostic information increased if the four lymphocyte tests were combined in a score (range 0-8). All 8 patients with pronounced lymphocyte defects (score 7-8), died, while all patients with score 0 and 1 were in complete remission. In contrast, total lymphocyte and T-lymphocyte counts and lymphocyte stimulation by PPD were of no prognostic value. The ability of certain lymphocyte functions to predict prognosis was equal or better than that of age and better than clinical staging, histopathology and symptoms.

The prognostic information of blood lymphocyte characteristics and clinical findings was assessed in 62 patients with chronic lymphocytic leukemia of B cell type. Bivariate and multivariate survival analyses were performed using age, Rai stage, surface membrane immunoglobulin (smIg) isotype pattern of the leukemic clone, total lymphocyte counts, numbers of proliferating lymphocytes and T cell subpopulations. Rai stages III and IV, high numbers of blood lymphocytes in S-phase (S+) and sm mu isotype were found to be partly independent factors predicting short therapy-free survival. Patients with a sm mu+ leukemic cell clone had a shorter therapy-free and total survival compared to those with sm mu+/delta+ and sm delta+ leukemic cells. Moreover, patients with high numbers of blood S+ lymphocytes had a shorter therapy-free and total survival compared to those with few S+ cells. These prognostic variables were valid also in patients with a low tumour burden (Rai stages 0, I and II) and may thus be of clinical importance as a guideline for therapeutic intervention.

The prevalence of complement dependent, cold-reactive lymphocytotoxic serum factors (LT) was studied in 80 untreated patients with Hodgkin's disease by a microcytotoxicity assay. Sera from 24 patients (30%) contained LT as judged from at least 50% lysis of lymphocytes from 16 to 23 randomly selected normal donors. The spontaneous incorporation of 14C-thymidine into blood lymphocytes from patients with LT was significantly higher than that of lymphocytes from LT-negative patients and from healthy controls. Total lymphocyte and T-cell counts, Concanavalin A, and pokeweed mitogen-induced lymphocyte DNA synthesis were lower in patients with LT. Lymphocytotoxic sera were more frequently encountered in patients with B symptoms, advanced disease, or nodular sclerosis/lymphocyte predominance histopathologic characteristics. LT were often found in patients with large and tumor-involved spleens. The ability of patient's serum to inhibit control lymphocyte response to Concanavalin A stimulation did not differ between LT-positive and LT-negative patients. We conclude that the presence of LT is associated with a quantitative and qualitative impairment of blood T-lymphocytes in untreated patients with Hodgkin's disease.

Spontaneous and mitogen induced blood lymphocyte DNA synthesis was studied in relation to weight and tumor involvement of the spleen in 33 untreated patients with Hodgkin's Disease. Splenic tumor involvement was found in 14 patients. In this group there was a strong inverse correlation between spleen weight and lymphocyte DNA synthesis induced by PWM, ConA and PPD. In patients with uninvolved spleens the spontaneous blood lymphocyte DNA synthesis increased with spleen weight. No correlation with the mitogen response was observed in this group.

Blood lymphocytes from twenty-three untreated patients with Hodgkin's disease and twelve healthy controls were studied for their ability to lyse tissue culture cells (Chang cells) labelled with [51Cr]chromate. Lysis induced by patients' lymphocytes in the presence of PHA or rabbit IgG antibodies to Chang cells (ACS) was impaired in some, and higher than normal in others. ACS-induced lysis showed some corlation with the content of lymphocytes carrying receptors for human complement (CRL) in the effectory population. No correlation with immunoglobulin-bearing cells was noted. PHA-induced cytoxicity did not correlate with lymphocyte subpopulations. The observations are consistent with the assumption that effector cells of antibody-induced lymphocyte-mediated cytotoxicity (K cells) may be present among CRL. The K-cell activity and PHA cytotoxicity by lymphocytes tended to decrease at high age, but impaired cytotoxity was also noted in young patients. A preliminary follow-up of patients 1-2 years after the beginning of treatment revealed almost abolished K-cell activity in four patients who died 6--13 months after testing. Patients in incomplete remission or with relapse after treatment had lower mean K-cell activity than those in complete remission. A similar, but less pronounced tendency was found for PHA-induced cytotoxicity. A prognostic role of impaired K-cell activity in Hodgkin's disease is suggested from these data, but requires confirmation in a larger clinical follow-up.

The surface membrane immunoglobulin (smIg) isotype pattern of the leukemic lymphocytes was studied in 66 unselected patients with CLL. Five distinct patient groups were identified according to the dominant heavy chain isotype(s); I: smmu + (n = 22), II: smmu +/smdelta + (n = 25), III: smdelta + (n = 4), IV: smgamma + (n = 5), V: no detectable heavy chains (n = 10). The majority of group I patients had a progressive disease at test while all patients in group II were in a non-progressive state. Moreover, the smIg pattern changed with the clinical activity of the disease: when the disease progressed, the relative number of smmu + cells increased and when patients entered an indolent stage after treatment the smmu + cell population decreased. In patients with stationary disease the smIg pattern remained essentially unchanged or the relative number of smdelta + cells increased. These observations might suggest that the smIg isotype pattern of the leukemic cell population reflects the biological behaviour and the clinical activity of the disease.

A Burkitt-like lymphoma/leukemia confined to bone marrow was detected in a human T cell leukemia virus (HTLV)-III/LAV- and Epstein-Barr virus (EBV)-seropositive homosexual man. The tumor cells were EBNA-positive and contained at least 22 EBV genomes per cell. They were totally immunoglobin negative, but showed other markers for B cells detected with monoclonal antibodies. The patient had an impaired cellular immunity to EBV antigens and EBV-infected cells at diagnosis, but these reactions normalized during treatment. Cell clones derived from the bone marrow tumor in vitro also carried EBV and had six different marker chromosomes, including the typical 14q+ chromosome and a t(8 - ;8), which resulted in trisomy for the largest part of 8q. Partial trisomy for 12q was also observed. The patient completed six courses of combination chemotherapy and remains in excellent health after 34 months of follow-up.

Two patients with Hodgkin's disease in remission and one chronic lymphatic leukemia patient with extraordinarily high anti-Epstein-Barr virus (EBV) (viral capsid antigen) antibody titers (greater than 10,000) were selected to study a spectrum of cell-mediated immune responses, including natural killer, interferon-boosted killer, antibody-dependent lymphocytotoxicity, and T-cell-mediated reactions. The purpose was to compare these reactions in patients with immunosuppression and a high EBV load who can hold their EBV-carrying cells under control with the corresponding reactions in patients with EBV-carrying lymphoproliferative disease. In contrast to the latter group, the three patients of the present study showed a less profound and less general suppression of the immune responses. Multiple effector mechanisms probably safeguard against the proliferation of EBV-transformed B-cells. Clinically manifest EBV-carrying lymphoproliferative disease occurs only in very severe immunodeficiencies effecting multiple effectors.

Sera from 296 unselected and untreated patients with non-Hodgkin lymphoma (NHL) classified according to the Rappaport and the Kiel systems were analyzed for antibodies to Epstein-Barr virus (EBV). The aim of the study was to determine whether antibody spectra and titers to EBV-coded antigens correlated to clinical and immunological variables and whether the titers were of any prognostic significance. Increased antibody titers to EB viral capsid antigen (VCA) and slightly raised titers to early antigens (EA) of the diffuse (D) and restricted (R) types were noted frequently. Anti-VCA antibody titers correlated to clinical stage and age of the patients but not to histological subgroups according to the Rappaport or the Kiel classification systems. However, anti-VCA titers greater than or equal to 1:2560 were seen only in diffuse lymphomas according to the Rappaport and in non-follicle cell-derived lymphomas according to the Kiel classifications. Patients with complement-receptor-positive diffuse lymphomas had higher anti-VCA titers than complement-receptor-positive nodular cases. Anti-VCA titers also correlated positively to serum IgG levels (p less than 0.01). Total number of lymphocytes separated from peripheral blood and mitogen induced (ConA, PWM) DNA synthesis were recorded before treatment in 56 of the patients. The patients exhibited a significant lymphocytopenia as well as a significantly reduced lymphocyte response to mitogens (p less than 0.001) compared to healthy controls. Elevated anti-VCA titers and anti-EA titers correlated to a good mitogen-induced lymphocyte response (p less than 0.05). Only anti-D 1:40 at diagnosis predicted a poor prognosis.

Spontaneously DNA synthesizing blood and spleen lymphocytes in 9 patients with Hodgkin's disease have been studied by surface markers and by autoradiography after in vitro labelling with 3H-thymidine of cell specimens separated according to size. The findings have been related to functional characteristics of blood lymphocytes. The distribution of E and EAC rosetting cells in blood and spleen was similar also after separation according to cell size. The number of mainly large activated blood and spleen cells was increased in patients as compared to controls. Both E and EAC rosetting cells participated in this unexplained cell challenge.

Blood lymphocytes from nineteen patients with Hodgkin's disease were tested in vitro before and after treatment. The patients were retested in complete and unmaintained remission at least 15 months after termination of radiotherapy. All patients except two had been treated with total nodal irradiation. The lymphocyte-DNA synthesis induced by concanavalin A (Con A) and PPD was poor and the spontaneous DNA synthesis was increased in untreated patients. Most patients had a T lymphocytopenia before treatment. After irradiation the total lymphocyte counts were reduced drastically. The number of T lymphocytes was particularly low, though the number of B lymphocytes decreased as well. However, the lymphocyte response to Con A and PPD remained low and unchanged. The results may suggest a persisting immunodeficiency in Hodgkin's disease as reflected by the lymphocyte response to Con A and PPD.

The study comprises 38 unselected and untreated patients with Hodgkin's disease (HD) and 23 healthy persons. Highly purified blood lymphocytes were analyzed for cells forming rosettes with sheep red blood cells (T lymphocytes), and lymphocytes bearing surface immunoglobulin (B lymphocytes) and/or carrying receptors for complement. Their DNA synthesis, spontaneously, or after activation with mitogens (phytohemagglutinin, concanavallin A, poke weed mitogen) and purified protein derivative (PPD) was measured. Delayed skin hypersensitivity to PPD and mumps antigen was studied. Most HD patients had low numbers of T lymphocytes (50% of the cases below normal range) while the mean B-lymphocyte level was normal but with a greater variation than in the control group. Lymphocytes from most patients were poorly stimulated by T-cell mitogens. Two-thirds of the patients and one healthy control had negative skin reaction to 2 TU PPD and the DNA synthesis of their lymphocytes after activation with PPD was low. Large lymphoid cells (greater than 9-mm diameter) were commonly present in HD blood and the spontaneous DNA synthesis was high, particularly in lymphocytes from stage B patients. The percentage of T lymphocytes and the stimulation of lymphocytes by T-cell mitogens or by PPD, a T-lymphocyte function, did not correlate and each test only detected defects in about half the cases. Simultaneous application of all tests revealed abnormalities of blood T lymphocytes in 33 out of 38 patients. Although the defects were usually more pronounced in patients with advanced disease, the impairment of T lymphocytes and their functions is present in all stages of Hodgkin's disease.

Monocyte depleted blood lymphocytes were analyzed in 28 patients with non-Hodgkin's lymphoma in early unmaintained complete clinical remission, in 8 long-term complete remission patients, and in 31 age-matched healthy controls, by SRBC rosetting and DNA synthesis induced by concanavalin A, pokeweed mitogen and PPD antigen. The median observation time from testing was 22 months (range 10-49 months). Early complete remission patients with a normal mitogen induced lymphocyte stimulation remained free of relapses. In contrast, patients who relapsed during the observation period had lymphocytopenia (p less than 0.05), reduced lymphocyte responses to mitogen (p less than 0.001) and to PPD antigen (p less than 0.05) as compared with healthy controls. Long-term complete remission patients did not differ from healthy controls with respect to lymphocyte counts and response to mitogen stimulation.

Purified peripheral blood lymphocytes (PBL) from nine untreated patients with Hodgkin's disease (HD), two HD patients in complete remission and 17 healthy donors were studied for natural killer (NK) cell activity against the K-562 cell line using a single cell cytotoxic assay, which allowed enumeration of effector cells and characterization of their surface membrane phenotypes alter staining with monoclonal antibodies. The frequency of NK ceils was significantly lower in HD patients than in controls (mean % ± s.d., 1.9 ± 0.9 and 2.8 ±1.2, respectively), while the fraction of target binding cells was similar in the two groups. The fraction of cytotoxic lymphocytes increased after pre-treatment of PBL with 500 iu leucocyte interferon in all tested control donors (n= 12) and the two patients in remission but only in four of seven untreated patients. No relation between the impaired NK cell frequency and age, tumour histology and clinical stage could be revealed, Subtyping of the target cell binding NK cells by monoclonal antibodies disclosed a marked heterogeneity of effector cells, NK effector cells reactive with Ml and anti-Ia antibodies were enriched while T3+, and T4+ NK lymphocytes tended to be reduced as compared to PBL, There was no difference between patients and healthy donors with regard to the surface antigen patterns of NK cells, Interferon treatment did not alter significantly the phenotypic characteristics of cytotoxic lymphocytes in patients and controls. It is concluded that the impairment of NK cell activity in HD is partly attributed to a lower frequency of cytotoxic effector cells among a normal number of target binding cells. The defect could not be attributed to a selective defect of effector cell subsets. [ABSTRACT FROM AUTHOR]