Immunological characterization of mononuclear cells in peripheral blood and regional lymph nodes of breast cancer patients.

Mononuclear cells from peripheral blood and draining lymph nodes of 40 patients with invasive locoregional breast cancer were examined for immunological cell surface markers (E, EAhuman, EAox, EAC, SIg pos.). Concomitantly, blood lymphocytes from 36 healthy women and axillary and mesenteric lymph-nodes from patients without malignant diseases were tested as controls. In peripheral blood of tumor patients E rosette-forming cells were slightly diminished as compared to the control group, whereas EAox and EAC rosette-forming cells were increased. These differences may be age-dependent rather than tumor-related. In the draining lymph nodes of breast cancer patients as well as in the control lymph nodes, the percentages of EAC rosette-forming cells and SIg positive lymphocytes were significantly increased compared to peripheral blood, whereas E and EAhuman rosette values remained unchanged. Percentages of EAox rosettes on the other hand were strongly diminished in the draining lymph nodes, suggesting that the EAhuman and EAox rosetting techniques detect 2 types of Fc-receptor bearing cells. No significant differences were found between the cell surface marker analysis of tumor-free and metastatic lymph nodes of breast cancer patients and the control lymph nodes.     

A thymus cell marker in murine leukemia virus-induced lymphomas of rats.

A specific marker for an immature population of thymus cells in the rat was shown by the rosette formation between thymus cells and guinea pig erythrocytes. This method was used to classify murine leukemia virus-induced rat lymphomas. Eight of nine Gross virus-induced rat lymphoma lines, which originated in the thymus, formed rosettes; whereas Friend, Rauscher, or Moloney virus-induced rat lymphoma lines, which originated in either the thymus, spleen, or mesenteric lymph nodes, did not form rosettes. The percentage of the total cells which formed rosettes in the Gross lymphoma lines decreased with in vivo passages. If the tumor cells were exposed to trypsin treatment, then the tumor cells would form rosettes. Lymphoma lines which lacked rosette-forming cells did not show rosette formation after trypsin treatment. An immunofluorescence test showed that none of the lymphoma lines induced by Gross, Friend, Rauscher, or Moloney viruses carried the surface immunoglobulin characteristic of B-cells. These results suggest that Gross lymphomas may be derived from the thymic cortex and that Friend, Rauscher, or Moloney lymphomas may be derived from either mature thymus cells (non-rosette-forming cells) or from a subpopulation of the B-cell series which does not have the surface immunoglobulin G receptor.     

Tumor-associated antigen in bovine and ovine lymphosarcoma.

Specific tumor-associated antigens were found on the membrane and in the cytoplasm of lymph node cells and peripheral blood lymphocytes (PBL) from cattle and sheep with lymphosarcoma by immunofluorescence tests. Materials from 15 cattle with the adult form of lymphosarcoma were examined. Cytoplasmic antigen was detected in fixed tumor cells from all 15 cases and in PBL from 9 cases tested. Membrane antigen was detected in living cells from 10 of the cases tested. In 3 calf-type cases, cytoplasmic antigen was found in a few (1 to 3%) of the tumor cells, while 1% of the cells from 2 thymic cases had cytoplasmic tumor antigen. In 15 cattle infected with bovine leukemia virus (BLV) but with no evidence of tumor, PBL from 3 cattle had the tumor-associated antigen in the cytoplasm. Negative results were obtained with similar tests done with 9 normal cattle that had no detectable BLV or BLV antibody. Cells from tumors induced with BLV in 5 sheep also had cytoplasmic antigen and membrane tumor-associated antigen. Tumor-associated antigen was found in PBL from 1 or 7 BLV-infected sheep with no clinical evidence of tumor. Similar tests were negative on 4 normal sheep.     

Concanavalin A and the production of bovine leukemia virus antigen in short-term lymphocyte cultures.

The influence of the mitogen concanavalin A (Con A) on the production of bovine leukemia virus (BLV) antigen in short-term lymphocyte cultures was determined by means of a single radial immunodiffusion test. Con A did not affect viral antigen production in peripheral blood lymphocytes from 60% of both experimentally and naturally infected cattle. Antigen production was stimulated by Con A in lymphocytes from 28% of the cattle, but it was inhibited in lymphocytes from 12%. Similar results were also obtained with lymphocytes from both blood and lymph nodes from 10 cattle with lymphosarcoma and from 10 clinically normal cattle with histologically normal lymph nodes. In sheep and goats, Con A had no effect on lymphocytes from 50%, stimulated BLV production in 43%, and inhibited BLV production in 7%. These results indicated that lymphocytes should be cultured with and without Con A to identify every BLV-infected animal.     

Subpopulations of human T-lymphocytes. XIX. T-cells and T-cells with receptors for IgMFc (T mu), IgGFc (T gamma), or IgAFc (T alpha) in the peripheral blood and regional lymph nodes of patients with untreated breast cancer

Peripheral blood and regional lymph node mononuclear cells from 43 untreated patients with breast cancer were analyzed for the proportions of total T-cells and T-cells with receptors for the Fc portion of IgM (T mu), IgG (T gamma), or IgA (T alpha). Proportions of total T-cells and T mu cells both in peripheral blood and lymph nodes from breast cancer patients were comparable to those from health controls. The proportion of T gamma cells, however, was significantly (P less than 0.01) increased in the peripheral blood and lymph nodes from breast cancer patients as compared to that from controls. The proportion of T alpha in the peripheral blood was comparable; however, when compared to the number of T alpha cells in control lymph nodes, T alpha cells were increased (P less than 0.025) in the regional lymph nodes from patients with breast cancer. When data on the proportions of T-cells and T-cell subsets were analyzed according to the presence or absence of metastatic disease in the regional lymph nodes, the proportion of T gamma cells was significantly (P less than 0.025) higher in the peripheral blood from patients with metastatic disease than in patients with nonmetastatic disease. This study demonstrates an abnormality of T-cell subsets in the peripheral blood and the regional lymph nodes from patients with breast cancer. The significance of these observations is discussed.     

Gamma-linolenic acid induces apoptosis in B-chronic lymphocytic leukaemia cells in vitro.

Gamma linolenic acid (GLA) is cytotoxic to many types of human cancer cells. Most chemotherapeutic agents are cytotoxic by inducing apoptosis. We examined the apoptotic activity of GLA on purified B-cells isolated from patients with B-cell chronic lymphocytic leukaemia (B-CLL) and from normal individuals. GLA significantly increased the degree of apoptosis in B-CLL B-cells after 24 hours of culture. The mean percentage of cells undergoing apoptosis when cultured in medium alone (spontaneous apoptosis) was 20% (range: 7 to 31%) (n=25) and in the presence of GLA (5 microg-60 microg) was: 42%-95%. In the presence of GLA 5 microg/ml and dexamethasone the degree of apoptosis was 86% (range: 72 to 100%). GLA induced apoptosis in B-CLL B-cells at a higher level than that observed with normal B-cells at all lower concentrations tested 5, 10 and 15 microg/ml: P=0.045; 0.027 and 0.022, respectively. At 30 microg/ml of GLA, no significant difference in the percentage of cells displaying apoptosis between B-CLL and normal B-cells was observed (P=0.075). GLA induced apoptosis in B-CLL T-cells at both 10 and 30 microg/ml. The degree of apoptosis in normal T-cells with GLA was also significant at the higher concentration of 30 microg/ml. Interleukin 4 (IL4), a viability factor in B-CLL, and vitamin E, an anti-oxidant, protected B-CLL B-cells against GLA (20 microg/ml)-induced apoptosis. These results demonstrate that GLA induces apoptosis in B-CLL B- and T-cells cells in vitro and that they are more susceptible to GLA-induced apoptosis than normal peripheral blood B- and T-cells.     

T and B lymphocytes in non-Hodgkin's lymphoma: a comparison of tumor-derived cells and peripheral blood lymphocytes.

T- and B-cell markers of lymphocytes in peripheral blood, involved node and spleen, PHA response of peripheral blood lymphocytes, serum immunoglobulin levels, and skin test reactivity to six common antigens were studied in 16 cases of untreated non-Hodgkin's lymphoma. Impaired response of peripheral lymphocytes to PHA was observed in 13 of 16 cases, regardless of the proportion of T lymphocytes. Of 12 cases in which skin tests were done, two were positive and had a normal PHA response, seven cases were positive in spite of low PHA response, and three were negative with low PHA response. In the lymph nodes from involved areas two cases showed monoclonal increase of B-cells, five showed "null" cell increase, and the remaining nine showed no increase or decrease of subpopulation of lymphocytes. No correlation with surface marker of lymphocytes to histologic classification was seen. From the above observations it was concluded: 1) a low PHA response in non-Hodgkin's lymphoma was not due to the decreased population of T-cells; 2) a low PHA response may not necessarily indicate impaired delayed hypersensitivity; and 3) non-Hodgkin's lymphoma can be classified in the following ways--B-cell proliferative type, "null" cell increase type, and normal T/B proportion type.     

Hand-mirror variant of acute lymphoblastic leukemia. Evidence for early T-cell lineage in two cases by evaluation with monoclonal antibodies

Lymphoid cells from two patients with hand-mirror variant of acute lymphoblastic leukemia (ALL) were studied with various monoclonal antibodies in attempts to determine their derivation and differentiation. The predominant feature of the malignant bone marrow cells was strong reactivity with antibody 3A1, which stains the majority of normal T-cells and is apparently present on all T-ALL cells. In addition, a less intense binding was observed with antibody 4F2, which reacts with activated or rapidly dividing cells, and antibody 10.2, which reacts with all thymocytes and most peripheral T-cells. Most other antibodies with a wide variety of specificities were not reactive or, in the case of a few anti-T-cell antibodies showed, by fluorescence-activated cell sorter analysis only weak staining on some cells. Sequential bone marrow studies in one patient, before and during treatment with chemotherapy, revealed a reduction of 3A1-positive cells, concordant with a decrease of malignant cells in the marrow. When involved lymph nodes, peripheral blood, or marrow were studied, similar reactivity patterns were found in all locations. The data obtained suggest that in both patients with hand-mirror variant of ALL, the malignant lymphoid cells were immature cells of early T-lymphocyte lineage. The relation of phenotype by monoclonal antibody analysis to hand-mirror morphologic type and biologic function is discussed.     

γ-Linolenic Acid Induces Apoptosis in B-Chronic Lymphocytic Leukaemia Cells in Vitro

Gamma linolenic acid (GLA) is cytotoxic to many types of human cancer cells. Most chemotherapeutic agents are cytotoxic by inducing apoptosis. We examined the apoptotic activity of GLA on purified B-cells isolated from patients with B-cell chronic lymphocytic leukaemia (B-CLL) and from normal individuals. GLA significantly increased the degree of apoptosis in B-CLL B-cells after 24 hours of culture. The mean percentage of cells undergoing apoptosis when cultured in medium alone (spontaneous apoptosis) was 20% (range: 7 to 31%) (n=25) and in the presence of GLA (5μg-60μg) was: 42%-95%. In the presence of GLA 5μg/ml and dexamethasone the degree of apoptosis was 86% (range: 72 to 100%). GLA induced apoptosis in B-CLL B-cells at a higher level than that observed with normal B-cells at all lower concentrations tested 5, 10 and 15μg/ml: P=0.045; 0.027 and 0.022, respectively. At 30μg/ml of GLA, no significant difference in the percentage of cells displaying apoptosis between B-CLL and normal B-cells was observed (P=0.075). GLA induced apoptosis in B-CLL T-cells at both 10 and 30μg/ml. The degree of apoptosis in normal T-cells with GLA was also significant at the higher concentration of 30μg/ml. Interleukin 4 (IL4), a viability factor in B-CLL, and vitamin E, an anti-oxidant, protected B-CLL B-cells against GLA (20μg/ml)-induced apoptosis. These results demonstrate that GLA induces apoptosis in B-CLL B-and T-cells cells in-vitro and that they are more susceptible to GLA-induced apoptosis than normal peripheral blood B-and T-cells.     

Goat lymphosarcoma from bovine leukemia virus

A goat given inoculations of sheep lymphocytes from cultures that produced bovine leukemia virus (BLV) died 8 years later with lymphosarcoma. The tumors were located in various lymph nodes, the mesentery, omentum, body wall, and retrobulbar tissues. The BLV had been cultured from lymphocytes during the first year after the goat's infection, and persisting BLV antibodies could be demonstrated when the animal was 7.5 years old. BLV provirus was identified by molecular hybridization in the DNA of the goat tumors at the above five locations. The tumors were similar to those found in lymphosarcoma of the adult bovine type (BLV associated). Normal goat liver, normal calf thymus, and calf-type lymphosarcoma (not BLV associated) served as negative controls. Our serologic, histologic, and molecular hybridization studies are evidence that the lymphosarcoma was induced BLV.     

Infiltrating lymphocytes in esophageal carcinoma--II. Subsets of T lymphocytes

The distribution of T lymphocytes and it's subsets in cancerous tissues, local lymph nodes and peripheral blood of patients with esophageal carcinoma were investigated. The percentages of total and active T cells in the peripheral blood and lymph nodes were within normal range. Most of the lymphocytes infiltrating in the cancerous tissues are T cells. It suggested that cellular immunity be developed in the cancerous tissues. The percentage of T mu in the patient's blood was lower than that of the normal person but the T gamma was higher. Proportions of T mu and T gamma in cancerous tissues were paralleled with those in blood, i.e. T gamma is higher than T mu. The mean percentage of T mu in lymph nodes was 25.52 +/- 14.06% and T gamma was 3.28 +/- 2.33%. It seems likely that the former is lower than normal and the latter is elevated.     

Lymphocyte response to pokeweed mitogen in chronic lymphocytic leukemia

The response of lymphocyte subpopulations to pokeweed mitogen (PWM) was studied in normal volunteers and patients with B-cell chronic lymphocytic leukemia (CLL). Since unfractionated peripheral blood mononuclear (PBM) cells from CLL patients consist of a markedly increased proportion of B-lymphocytes and a decreased proportion of T-lymphocytes, enriched fractions of CLL B-cells and CLL T-cells were cultured in 1:1 proportions in autologous and allogeneic combinations with normal B-cell and T-cell-enriched fractions. Cultures containing normal B-cells with either autologous or allogeneic normal T-cells responded well to PWM. CLL T-cells were capable of providing a helper function for both proliferation and differentiation of normal B-cells, which was not significantly different from that provided by allogeneic normal T-cells. CLL B-lymphocytes were unresponsive to PWM when cultured in the presence of either autologous CLL T-lymphocytes or allogeneic normal T-lymphocytes. The responsiveness of CLL B-cells was not restored by the addition of normal peripheral blood monocytes to the cultures. These experiments indicate that there is an intrinsic B-cell defect which prevents CLL B-lymphocytes from responding to PWM.     

Canine transmissible venereal sarcoma: distribution of T and B lymphocytes in blood, draining lymph nodes and tumours at different stages of growth.

The levels of T, B and null lymphocytes in the peripheral blood, draining lymph nodes, and tumour masses at different growth stages in dogs transplanted with canine transmissible venereal sarcoma (CTVS) were determined by immunofluorescence techniques. The tumours were classified at excision into "progressor", "steady state", and "regressor" stages of growth. The percentage of B cells in the lymphocytes infiltrating into the progressively growing tumours (n = 10, 37.3 +/- 7.4%) was significantly higher (P less than 0.025) than that in regressing tumours (n = 21, 26.1 +/- 1.9%). In contrast, the percentage of T cells in the lymphocytes infiltrating into the regressing tumours (n = 21, 61.2 +/- 2.6%) was significantly higher (P less than 0.005) than that in the progressively growing tumours (n = 10, 34.0 +/- 5.1%). The tumours at the steady-state growth stage (n = 9) had 50.8 +/- 5.7% infiltrating T-cells, which was significantly higher (P less than 0.005) than the progressors and lower (P less than 0.005) than the regressors. The percentage of null cells of progressors (n = 10, 26.0 +/- 6.9%) was significantly (P less than 0.025) higher than in regressors (n = 21, 13.5 +/- 2.9%). The draining lymph nodes of progressor dogs (n = 5) had significantly fewer (P less than 0.025) B cells (8.2 +/- 2.3%) than in normal (n = 5, 16.1 +/- 3.1%), regressors (n = 12, 19.1 +/- 1.7%) and steady-state dogs (n = 5, 15.8 +/- 2.6%). Although there was slight lymphopenia and fluctuation of null cells, no significant differences in T- and B-lymphocyte levels were noted in the peripheral blood of the tumour dogs (n = 44) studied.     

Studies on the use of membrane markers for the differential diagnosis of malignant lymphomas.

Lymphoid cells isolated from lymph nodes, spleen and the peripheral blood were examined using the rosette test with sheep erythrocytes (E), immunofluorescent staining of the surface immunoglobulins (SmIg) and the combined test (SmIg + E). The studies were performed on 48 patients with untreated non-Hodgkin lymphoma, 14 Hodgkin patients, 5 patients with lymphadenitis and on 132 controls. In the control group the following percentages of B-T lymphocytes were obtained: blood (n = 100); T-68 +/- 9, B-20 +/- 6, 0-11 +/- 7, BT-1 +/- 1, lymph nodes (n = 24): T-62 +/- 13, B-19 +/- 11, 0-18 +/- 8, BT-1 +/- 1, spleens (n = 8): T-45 +/- 9, B-33 +/- 9, 0-23 +/- 7, BT-1 +/- 1. In non-Hodgkin lymphoma lymph nodes a statistically significant increase of B and 'null' cells was noted in comparison to controls and to the Hodgkin lymphoma group. The characteristics of the Hodgkin nodes did not differ significantly from controls. After arranging the non-Hodgkin lymphomas according to the Kiel classification it was observed that the low-grade malignant lymphomas subtype most often an immunological 'B type' and 'mixed B/0' type was present whereas the high-grade malignant lymphoma group showed a distinct majority of 'null' cells. It was concluded that the surface markers studied allow a differentiation in most cases between the non-Hodgkin lymphomas and other disorders of the lymphoid system. They are not sufficient to distinguish the subtypes of non-Hodgkin lymphomas but do give interesting information about the more exact nature of the disorder.     

胃癌患者不同组织CD44v5+细胞表达率的比较

 目的　探讨胃癌患者不同组织CD4 4v5的表达规律。方法　采用流式细胞术对于 39例胃癌患者的癌灶、癌旁组织、淋巴结转移灶和外周血细胞的CD4 4v5表达率进行了检测和对比分析。结果　胃癌患者不同组织的CD4 4v5 +细胞检出率均显著高于正常对照组 (P <0 .0 5或P <0 .0 1)。胃癌患者各种组织CD4 4v5 +细胞表达率大小顺序为癌灶 >癌旁组织 >淋巴结转移灶 >外周血细胞 ,前三者和外周血细胞之间有显著性差异 (P <0 .0 5或P <0 .0 1) ,而前三者之间却无显著性差异 (P >0 .0 5 )。在胃癌患者的各种组织中 ,伴肿瘤转移患者的CD4 4v5 +细胞检出率均显著高于未转移者 (P <0 .0 5或P <0 .0 1)。结论　胃癌患者身体各种组织的CD4 4v5 +细胞表达率显著升高 ,且CD4 4v5 +细胞表达率与肿瘤转移密切相关。     

Relationship between chronic lymphocytic leukemia and small lymphocytic lymphoma. A comparative study of membrane phenotypes in 270 cases.

BACKGROUND. Chronic lymphocytic leukemia (CLL) and small lymphocytic lymphoma (SLL) are considered different tissue expressions of the same disease process, although they are clinically separable nosologic entities. A systematic comparison of the membrane phenotypes in the two entities needs to be investigated. METHODS. Cell suspensions from peripheral blood of 184 patients with CLL, bone marrow from 23 patients with CLL, and lymph nodes from 86 patients with SLL were analyzed to compare the membrane phenotypes. RESULTS. There were no significant differences between the three groups in the mean percentages of cells expressing surface immunoglobulin (SIg), CD5, CD19, CD20, and CD2 or in the frequency of cases with weak SIg. Although the mean percentage of mouse rosette-forming cells (MRFC) showed no statistical difference between bone marrow from CLL and lymph nodes from SLL, the mean percentage of MRFC in peripheral blood from CLL (48.02 +/- 18.23%) was twice as high as that in bone marrow from CLL (25.27 +/- 21.51%) and lymph nodes from SLL (20.87 +/- 16.72%) (P less than 0.001). Correlation analysis assessing the association of MRFC and residual T-cells showed a negative coefficient (r), and the r was statistically significant in bone marrow from CLL and lymph nodes from SLL but not in peripheral blood from CLL. The mean CD4/CD8 ratios in descending order were as follows: the ratio in lymph nodes from SLL (4.25) was greater than that in peripheral blood from CLL (1.70), which was greater than that in bone marrow from CLL (0.82); this followed the same pattern as the respective tissue controls. The mean ratios were not statistically different from those of their respective control groups. CONCLUSIONS. The similarity of membrane phenotypes between CLL and SLL provided evidence that the two are different tissue expressions of the same disease. The alterations in CD4/CD8 ratios were related to the type of tissue analyzed and not to the disease process. The difference in MRFC presumably results from the microenvironment or residual T-cells.     

Immunoblastic Lymphadenopahy in a Five-month-old Girl: Successful Treatment with Immunosuppressants

A five-month-old girl developed high fever, erythema, hepatosplenomegalyand generalized lymphadenopathy. Laboratory examinations revealedelevated peripheral leukocyte counts, C-reactive protein, lactatedehydrogenase and serum ferritin level. Pathologic examinationof the lymph nodes revealed immunoblastic lymphadenopathy (IBL)on the basis of the complete effacement of the normal architecture,replacement by a diffuse infiltrate composed of immunoblasts,plasmacytoid cells and small lymphocytes, and an abortive proliferationof blood vessels. B-cells and T-cells were nearly equally mixedthroughout the lymph nodes. No rearrangements of the Bcell immunoglobulinand T-cell receptor genes were detected. The patient was initiallytreated with -interferon with dramatic efficacy. After relapse,however, the disease was well controlled with cyclosporin A(CyA) and subsequently with combination regimens of CyA, deoxyspagarinand azathioprine with fair success. An alternating regimen of6-mercaptopurine, cyclophosphamide and methotrexate was theninstituted which continued the complete remission for 12 months.The effects of immunosuppressants in the treatment of IBL meritinvestigation.     

Growth and spread in nude mice of Epstein-Barr virus transformed B-cells from a chronic lymphocytic leukemia patient

Subcutaneous inoculation of Epstein-Barr virus (EBV) transformed peripheral blood B-lymphocytes (PBL) from an untreated chronic lymphocytic leukemia (CLL) patient produced progressively growing lethal tumors in 4 of 11 whole body irradiated (440 rads) nude mice. In one tumor bearing mouse there was splenomegaly and generalized enlargement of lymph nodes. Chromosomal analysis and membrane immunofluorescence revealed that cells in all the 4 s.c. tumors and a proportion of cells in the enlarged spleen and lymph nodes had human chromosomes and contained human kappa or lambda chains demonstrating that these were polyclonal human B-cells. Epstein-Barr virus associated nuclear antigen could be detected in 100% of cells in all the 4 EBV transformed B-cell lines in vitro and aliquots of cells from several s.c. tumors and metastatic lesions examined. Successful serial transplantation into irradiated nude mice was possible for at least 3 generations with one of the 4 s.c. tumors. During serial transplantation, spread of tumor cells to the spleen and lymph nodes could be detected in all the 3 passage mice investigated; however, there was no evidence in any mouse of dissemination of tumor cells into the bloodstream or into any organ other than lymph nodes and spleen. s.c. tumors also developed in a proportion of irradiated nude mice after inoculation of cells from two other s.c. tumors and the metastatic spleen and lymph nodes, but all these tumors regressed during the first or second transplant passage. Two % of PBL from the untreated patient and 4% of EBV transformed PBL maintained in vitro were found to have trisomy of chromosome 12 which is the most frequently reported anomaly associated with human CLL B-cells. It is highly probable that the cells with trisomy were derived from the leukemic clone of this patient. Cells with this trisomy predominated in most metastatic sites compared to the parent s.c. tumors. Inoculation of irradiated nude mice with EBV transformed PBL from this patient after chlorambucil therapy (100% metaphase plates with 46,XY,11q+ karyotype) or with EBV transformed PBL from 2 normal adults failed to produce any progressively growing tumor in a total of 12 irradiated animals observed greater than 300 days. Although there are several reports of EBV induced immortalization of CLL B-cells in vitro, we have not seen any previous report on the successful serial transplantation and dissemination of EBV transformed CLL B-cells in nude mice.     

Differentiation between benign and malignant human lymph nodes by means of immunologic markers.

A surface-marker assay combining immunofluorescence with anti-human immunoglobulin or anti-human brain serum (AHBS) and the formation of rosettes with untreated (E), antibody-sensitized (EA) and complement-coated (EAC) sheep erythrocytes was used to study mononuclear cell suspensions of human lymph nodes. The frequency of cells expressing more than one marker was increased in lymphoma nodes as compared to normal and hyperplastic nodes. The cells which simultaneously expressed complement receptors, surface immunoglobulin and the marker identified by AHBS represented the most prominent and characteristic subpopulation identified in neoplastic nodes. Distributions of cells with double and triple markers were studied by combining immunofluorescence with rosetting on frozen tissue sections. The multiple-marker cells had distributions that were characteristic in different human lymphomas. Benign and malignant human nodes could be distinguished on the basis of frequency and distribution of mononuclear cell populations carrying distinctive combinations of T- and B-cell surface markers.     

Increase in the proportion of cells with the C'3 receptor in Balb/c mice bearing mammary tumors

Transplantation of mammary tumors originally induced by dimethylbenzanthracene caused splenomegaly and an increase in the total population of spleen cells paralleling the increase in tumor size. At least part of the hypercellularity was due to in situ proliferation as evidenced by the increased number of blast forms and mitotic figures. The cells were characterized with respect to parameters generally associated with B-cells; surface immunoglobulins as detected by immunofluorescence and C'3 receptors as determined by rosette formation with sheep red blood cells (SRBC) coated with rabbit anti-SRBC and mouse complement. There was no change in the percentage of B-cells with surface immunoglobulin in tumor-bearing mice compared with the control animals. However, there was a profound change in the representation of cells bearing C'3 receptors. The percentage of the cells increased dramatically with tumor growth. Various possibilities regarding the nature and function of these cells are considered.