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PubMed1987

The ontogeny of germinal centre forming capacity of neonatal rat spleen.

In non-specifically immunized rats, bred under conventional conditions, the first 'spontaneous' germinal centres were observed by 21 days after birth. Deliberate antigenic stimulation led to an earlier appearance of germinal centres in neonatal spleen: immunization with sheep red blood cells as early as 7 days after birth resulted in germinal centre formation in the spleen as observed 7 days later. By that time the first primary follicles could also be observed, in both immunized and non-immunized rats. Although 3-day-old rats upon antigenic stimulation failed to generate germinal centres in their spleen, transfer experiments of 3-day-old spleen cells to lethally X-irradiated syngeneic adult recipients indicated that 3-day-old spleens at least contained all the essential lymphoid elements (B and T cells) needed for germinal centre formation. These results strongly suggest that the failure to induce germinal centres in 3-day-old rats is most probably due to an immaturity of their splenic microenvironment. Immunohistochemical staining of frozen sections of neonatal rat spleen using mAb ED 5 and MRC OX-2 showed that follicular dendritic cells (FDC) were found as soon as primary follicles were found (i.e. by 14 days after birth). The appearance of FDC in neonatal spleens was not influenced by deliberate antigenic stimulation nor by the administration of adult spleen cells. We postulate that, during the development of FDC, a splenic microenvironment is created that allows primary follicle formation and the generation of germinal centres.

Animals↗
PubMed1987

Effect of plasmacytoma cells on the production of granulocyte-macrophage colony-stimulating activity (GM-CSA) in the spleen of tumor-bearing mice.

Mice bearing syngeneic plasma cell tumors are characterized by elevated numbers of granulocyte-macrophage progenitors (GM-CFU) in the spleen. We investigated the role of syngeneic plasmacytomas in the hematologic response to tumor cell transplantation by assaying the production of granulocyte-macrophage colony-stimulating activity (GM-CSA) by cultured spleen cells of tumor-bearing mice and by plasmacytoma cells, alone and in coculture with spleen cells. Elevated levels of GM-CSA were detected in 7-day culture supernatants of spleen cells from Balb/c mice transplanted 2 weeks previously with syngeneic 4T00.1 plasmacytoma cells. Colony assays of spleen cells from tumor-bearing mice demonstrated the presence of both granulocyte-macrophage and tumor cell colonies. A high frequency of GM-CFU was detected in cultures which had not been supplemented with an exogenous source of GM-CSA. Significant levels of GM-CSA were detected in media conditioned by 4T00.1 plasmacytoma cells. 4T00.1-conditioned medium did not stimulate the growth of primative erythroid (BFU-E) and multilineage (CFU-GEMM) colonies, but stimulated the growth of FDC-P1 cells, thereby establishing the activity produced by 4T00.1 cells as GM-CSF. The levels of GM-CSA in media conditioned by coculturing control spleen and 4T00.1 cells were significantly higher than those detected in media conditioned by spleen cells alone. The colony frequency induced by the coculture supernatants, however, did not exceed the sum of the colonies detected in marrow cell cultures stimulated with media conditioned by control spleen and 4T00.1 cells alone. Our findings demonstrate that murine plasmacytoma cells are capable of secreting GM-CSF. They further suggest a key role for GM-CSA production by tumor cells in the hemopoietic response of mice bearing syngeneic plasma cell tumors.

Animals↗
PubMed1986

Characteristics of hemopoietic stem cells determined by single spleen colony transplantation.

By using a spleen colony transfer technique and sex chromosome determinations as a cytogenetic marker, we compared the spleen colony-forming cells of bone marrow origin from three strains of mice, i.e., LACA, C57, and (LACA female X C57 male) F1. The proliferation potential of spleen colony-forming cells was found to be heterogeneous in nature. Some of the cells have the essential characteristics of hemopoietic stem cells (Ps-CFUS), i.e., the capacity for sustained proliferation and differentiation into myeloid and lymphoid lineages of cells, whereas others exhibit the capacity to form gross colonies on irradiated recipient spleen which are capable of differentiating different lineages of blood cells, but have lost the ability to reconstitute hemopoiesis in lethally irradiated recipients (Pg-CFUS). The results of studies on spleen colony-forming cells of F1 mice indicate the existence of a subpopulation of primitive precursors of spleen colony-forming cells (Pre-CFUs) which are not able to form spleen colonies on recipient spleens, but, under appropriate stimuli, may assume the properties of Ps-CFUS. Therefore the sequential process of blood cell differentiation may be described as follows: Pre-CFUS----Ps-CFUS----Pg-CFUS----different lineages of hemopoietic progenitors----different lineages of blood cells.

Animals↗
PubMed1979

In vivo activity of spleen cells from untreated syngeneic mice against Ehrlich ascites carcinoma.

Inbred CFW/L1 or BALB/c male mice were injected intraperitoneally with 1 X 10(5) EAC cells. They died in 98.6 or 100%, respectively, within 100 days of observation. Intraperitoneal injection of 6 X 10(7) syngeneic spleen cells from 6--9-week-old CFW/L1 or BALB/c donors 2.5--4 h after tumour inoculation significantly prolonged MST and 16.7 or 23.5% of mice, respectively, survived for 100 days. Smaller amounts of spleen cells, thymus cells, mixed spleen and thymus cells, or spleen cells from older donors did not show this effect. Homogenized spleen cells were also inactive. Antitumour activity of spleen cells was still evident when they were isolated from irradiated (2 000R) donors or were given 24 h after tumour inoculation. Spleen cells used for in vivo studies showed spontaneous cytotoxic activity in vitro against EAC cells in the 51Cr-release cell-mediated cytotoxicity test. The possible participation of NK cells in the observed in vivo antitumour activity of normal spleen cells is discussed.

Animals↗
PubMed1979

Spleen lipids: effect of whole body gamma irradiation and radioprotective chemicals.

Effect of whole body gamma irradiation (1200 r) on spleen lipid metabolism of male and female rats 24 hrs and 48 hrs after irradiation and the effect of radioprotective chemicals vis. AET, serotonin, their mixture and cystamine on radiation induced changes in spleen lipid metabolism has been studied. In male rats both 24 and 48 hrs after irradiation a significant increase in spleen total lipids, cholesterol, phospholipids, phosphatidylethanolamine and phosphatidylcholine was observed. Administration of AET before irradiation prevented the changes in spleen total lipids and cholesterol but not in phospholipids, which was prevented by prior administration of both serotonin and the mixture of serotonin and AET. In female rats 24 hrs after irradiation only spleen total lipids showed an increase which was prevented by prior administration of cystamine. In male rats, 24 hrs after irradiation the incorporation of NaH2 32PO4 (counts/min/ug PLP and counts/min/g spleen) into spleen total phospholipids, phosphatidylcholine and phosphatidylethanolamine was reduced and this was corrected by prior administration of AET. Serotonin and the mixture of serotonin + AET did not protect the specific activity of phosphatidy choline. In female rats irradiation increased the incorporation of NaH2 32PO4 into phosphatidylcholine, which was not prevented by prior administration of cystamine. The fatty acid composition of spleen lipid of female rats was profoundly altered 24 hrs after irradiation. Palmitic acid and oleic acid showed an increase whereas arachidonic and fatty acid above arachidonic acid showed an decrease, which were corrected by administration of cystamine before irradiation.

Animals↗
PubMed1982

The sequestration capacity of "hypersplenic" rat spleens for heat-damaged erythrocytes.

The sequestration capacity of the spleens of controls and hypersplenic rats was examined. Hypersplenism was induced by long-term intraperitoneal application of methyl cellulose. The animals were injected single doses of various amounts of heat-damaged 51Cr-labelled erythrocytes (0.1 ml to 1.5 ml); radioactivity in spleen was determined 4 hrs. following application. The amount of red cells sequestrated in the spleens of hypersplenic animals was significantly increased against the controls, after administration of massive volumes of cells. The maximum amount of erythrocytes sequestrated in the spleens of the control rats amounted to an average weight of spleens 1.25 g to 0.158 ml (0.126 ml per g of spleen), and in hypersplenic animals to an average weight of the spleen of 4.87 g to 0.283 ml (0.058 ml/l g of spleen weight).

Animals↗
PubMed1984

Cyclophosphamide-induced appearance of immunopotentiating T-cells in the spleens of mice bearing a large MOPC-315 tumor.

Following low-dose cyclophosphamide (CY) therapy (15 mg/kg) of mice bearing a large MOPC-315 tumor, the suppressive activity of their Sephadex G-10-adherent spleen cells (primarily macrophages) is overcome. Accordingly, when Sephadex G-10-adherent spleen cells from CY-treated tumor-bearing mice are added to the in vitro immunization culture of normal spleen cells, they do not suppress but actually bring about the generation of an augmented level of antitumor cytotoxicity. The ability to enhance the generation of antitumor cytotoxicity appears in the Sephadex G-10-adherent spleen cell population by Day 5 post-CY therapy of tumor-bearing mice and persists for at least 55 days; no such immunopotentiation is observed following administration of a low dose of CY to normal mice. In order for the immunopotentiating cells from CY-treated tumor-bearing mice to be effective in enhancing the generation of antitumor cytotoxicity, they must be added to the immunization culture of normal spleen cells no later than Day 3 (out of the 5 days) post-culture initiation. The CY-induced immunopotentiating activity resides in the T-cells, as is evident from the following observations. The immunopotentiating activity was abolished when the Sephadex G-10-adherent spleen cell population from CY-treated tumor-bearing mice was depleted of T-cells by anti-Thy 1.2 plus complement but not when this adherent spleen cell population was depleted of macrophages by carbonyl iron and magnet. Moreover, the immunopotentiating activity was also present in a population of CY-treated tumor-bearer spleen cells highly enriched for T-cells by passage through nylon wool columns. Thus, low-dose CY therapy overcomes the immunosuppressive activity of macrophages and induces the appearance of T-cell-mediated immunopotentiating activity, thereby leading to the development of an augmented level of antitumor cytotoxicity that can cooperate effectively with the tumoricidal activity of CY in the eradication of a late-stage, large s.c. tumor and extensive metastases.

Adjuvants, Immunologic↗
PubMed1983

Proliferation and emigration of newly formed lymphocytes from pig spleens during an immune response.

Normal young pigs were immunized intravenously with sheep red blood cells (SRBC). At various times after a second SRBC injection the spleens were connected to an extracorporeal perfusion system, and proliferating lymphoid cells in the spleens were selectively labelled with tritiated thymidine. One day later the relative and absolute numbers of spleen-derived lymphocytes were determined by autoradiography in the following organs: various parts of the spleen, mesenteric and cervical lymph nodes, thymus, bone marrow, Peyer's patches, tonsils, intestine, lung, liver and blood. From 1 to 7 days after the second SRBC injection, the spleens produced increasing numbers of lymphocytes, and labelled cells were found especially in the blood and bone marrow. The newly formed splenic lymphocytes migrated preferentially to T- but also to B-cell areas in lymph nodes, Peyer's patches and tonsils. In all organs outside the spleen nearly all labelled spleen-derived lymphocytes were small lymphocytes. However, the bone marrow contained a high proportion of labelled immature and mature plasma cells. The spleen produced large numbers of lymphocytes during the secondary immune response, many of which migrated to different organs probably as memory cells, while others were found in the bone marrow as effector cells from the immune response.

Animals↗
PubMed1984

[Experimental studies on the rat-model for partial splenectomy and on spleen regeneration].

Partial splenectomy is often preferred to total splenectomy as a method to prevent serious post-operative sepsis. However, most of the previous researches on spleen function following partial splenectomy have failed to provide accurate data on the minimum increase of residual spleen in weight necessary for protection against postoperative sepsis. This is not only to the lack of appropriate model but also to the negligence of the effects of spleen regeneration after partial splenectomy. To produce an accurate model for partial splenectomies, and to know how the spleen regenerates following partial resection, about 500 rats underwent partial splenectomies. From these experiments, the following results were obtained. These partial splenectomies based on length of the spleen were satisfactory models as partial splenectomy. Absolute volume of regenerated spleen was finally almost the same in every model. However, the less the remnant spleen was the longer period was necessary until the spleen regeneration completed.

Animals↗
PubMed1982

Mode of action of polyethylene glycol 6000 in potentiating the in vitro generation of antitumor cytotoxicity by MOPC-315 tumor bearer spleen cells.

Some of the possible mechanisms by which polyethylene glycol (PEG) augments the ability of MOPC-315 tumor bearer spleen cells to mediate in vitro antitumor cytotoxicity were evaluated. The level of antitumor cytotoxicity obtained in 5-day cultures of tumor bearer spleen cell suspensions correlated inversely with the percentage of Trinitrophenol (TNP)-rosettable cells (presumably metastatic tumor cells) present in the spleen. The kinetics of decrease in the percentage of TNP-rosettable cells coincided with the appearance of antitumor cytotoxicity. In addition, PEG was shown to interfere with the ability of viable tumor cells to suppress the in vitro generation of antitumor cytotoxicity in normal spleen cells cultured with mitomycin C-treated tumor cells. However, the decrease in the content of TNP-rosettable cells and the concurrent increase in the level of antitumor cytotoxicity were not due to direct cytotoxic and/or cytostatic effects of PEG on tumor cells. Spleen cells cultured in the presence of PEG had an increased rate of [3H]thymidine incorporation and proliferation compared to spleen cells cultured in the absence of PEG. However, the PEG-induced decrease in the percentage of TNP-rosettable cells either preceded or occurred at the same time that the PEG-induced increase in spleen cell number was observed. Therefore, spleen cell proliferation can at best explain only partially the PEG-induced decrease in the content of TNP-rosettable cells, and other mechanisms for the decrease must be considered.

Animals↗
PubMed1980

Production of cytotoxic factor in the spleen of dengue virus-infected mice.

Swiss albino mice inoculated i.c. with dengue type 2 virus (DV) show presence of a factor in the spleen which is cytotoxic to the normal mice spleen cells in vitro. The cytotoxic factor is present in the homogenate as well as in the culture supernatants of the spleen cells. Maximum cytotoxic activity is present in the spleen of moribund mice on 10-11 p.i. day. No cytotoxic activity was found in the homogenates of brain, liver, heart and skeletal muscles. Sera from a few mice only had cytotoxic activity and were not complement-dependent. Cytotoxic activity in the homogenate and culture supernatant (TCF) of various spleen cell populations was screened. Cytotoxicity of glass-non-adherent cells was significantly higher. DV-infected spleen cells, depleted of macrophages by carbonyl iron treatment, had higher cytotoxic activity. B-cell-enriched fractions and the glass-adherent cells had negligible cytotoxic activity. Purified lymphocytes and the T-lymphocyte-enriched fractions had maximum cytotoxicity. Thus, a cytotoxic factor is produced by T lymphocytes of the spleen of DV-infected mice which kills normal mouse spleen cells in vitro.

Animals↗
PubMed1980

Inhibition of B-lymphocyte clonal proliferation by spleen cells from plasmacytoma-bearing mice.

This study demonstrated that spleen cells of inbred BALB/c mice with plasmacytomas inhibited proliferation of the B-lymphocyte colony-forming cells that were precursors of immunoglobulin-producing cells. The number of B-lymphocyte colonies/10(5) nucleated spleen cells was depressed in mice with plasmacytomas. The degree of suppression correlated with the size of the tumor. Spleen cells from tumor-bearing mice suppressed the formation of B-lymphocyte colonies by normal BALB/c spleen cells. Treatment of the myeloma spleen cell suspensions with anti-immunoglobulins, anti-theta, or 1,500 rads did not prevent the suppression. Spleen cells from tumor-bearing mice depleted of adherent cells by passage over Sephadex G-10 columns or adherence to plastic dishes did not retain their suppressive activity. Adherent spleen cells inhibited B-lymphocyte colony formation when they were either in direct contact with normal spleen cells or separated from them by a layer of agar. These studies suggested that the immune suppression seen in plasmacytoma-bearing mice occurred partially at the level of the B-lymphocyte progenitor cell and that this suppression was mediated by an adherent mononuclear cell.

Animals↗
PubMed1981

Metastatic properties conferred on nonmetastatic tumors by hybridization of spleen B-lymphocytes with plasmacytoma cells.

The P3-NSI/1-Ag4-1 (NSI) plasmacytoma, when transplanted sc in syngeneic BALB/c mice, grows locally without forming spontaneous metastases. We tested whether somatic hybridization of the NSI cells with spleen B-lymphocytes would render them metastatic in (C57BL/6 X BALB/c) F1 mice. We found that the hybridomas thus produced generated spontaneous metastases with distinct organ specificities. Some hybridomas produced metastases in both liver and spleen, whereas others produced metastases in only the liver. Cells derived from spleen-and liver-seeking hybridomas, when transplanted sc, produced tumors that metastasized to both the liver and spleen. Tumor cells derived from spleen metastasis produced, on transplantation, a tumor that generated spleen metastasis of a larger mass than did tumors derived from liver metastases. Cells derived from liver-seeking hybridomas metastasized to only the liver. Similar patterns of organ specificity were observed after iv injection of the hybridoma cells. The spleen seemed to play determining role in controlling the production of liver metastases by hybridomas that produced both liver and spleen metastases. Such hybridomas did not produce liver metastases when injected into splenectomized recipients. Hybridomas that were only liver-seeking did produce metastases in splenectomized recipients.

Animals↗
PubMed1980

Effect of polyethylene glycol 6000 on the generation of antitumor cytotoxicity in MOPC-315 tumor bearer spleen cells cultured in the presence or absence of inactivated stimulator tumor cells.

Noncytotoxic spleen cells from BALB/c mice bearing 15- to 26-mm (but not 29-mm) s.c. MOPC-315 tumors that were cultured in medium containing 2% polyethylene glycol 6000 (PEG) developed substantial levels of anti-MOPC-315 cytotoxicity as assayed by 51Cr release. The level of cytotoxicity obtained increased with progression of tumor growth. Addition of mitomycin C-treated stimulator tumor cells and PEG to the culture of tumor bearer spleen cells resulted in augmentation of antitumor cytotoxicity to a level that was greater than the sum of the levels of cytotoxicity exhibited by spleen cells cultured in the presence of either mitomycin C-treated tumor cells or PEG. Maximal levels developed when the spleen cells were cultured for 5 to 6 days in 2% PEG at a responder/stimulator cell ratio of 15/1 or 30/1. Tumor bearer spleen cells that were cultured in PEG with or without added MOPC-315 stimulator cells exhibited strong anti-MOPC-315 cytotoxicity but were virtually noncytotoxic to allogeneic EL4 and syngeneic blast cells. Furthermore, these spleen cells were far superior to spleen cells cultured without PEG in mediating in in vivo antitumor activity in the local adoptive transfer assay. Thus, tumor bearer spleen cells cultured in the presence of PEG might be useful in immunotherapeutic regimens requiring histocompatible cells with augmented antitumor cytotoxicity but devoid of reactivity against normal cells.

Animals↗
PubMed1997

Sarcoid-like reaction in the regional lymph nodes and spleen in gastric carcinoma: a clinicopathologic study of five cases.

To clarify the occurrence of sarcoid-like reaction in the spleen of the gastric carcinoma patients, 100 consecutive specimens from gastrosplenectomy were examined. Sarcoid-like reaction was observed in the lymph nodes of 13 cases (13%) and the spleen of five cases (5%). All cases of the latter group were included in the former one. None of them showed any symptoms or signs indicative of systemic sarcoidosis. It seems that the cases with sarcoid-like reaction in the spleen ocurred more frequently in an advanced stage of the gastric cancer than those without this phenomenon. Epithelioid cell granulomas (EPGs) appeared to arise in the periarteriolar lymphoid sheaths of the spleen histologically, but were never found in red pulp or germinal centers. They were composed of groups of epithelioid cells and accompanied by the small lymphocytes and plasma cells. In three cases, scattered eosinophils were also observed among the epithelioid cells. Immunohistochemically, the majority of the intragranulomatous small lymphocytes had T-cell phenotype, while B-cells formed only the minor cellular population. None of the 13 cases contained EPGs in the primary tumor. Our study indicates that sarcoid-like reaction in the spleen is possibly not such a rare phenomenon in the gastric cancer as previously considered and more frequently seen in the advanced stage of the gastric cancer. Sarcoid-like reactions of the regional lymph nodes were more frequently seen in the patients with EPGs in the spleen than in those without. We also suggest that the incidence of sarcoid-like reactions in the spleen is closely related to those in pancreaticosplenic nodes and/or nodes of the hilus of the spleen.

Aged↗
PubMed1997

Spleen-derived stromal cells. Adhesion molecules expression and lymphocyte adhesion to reticular cells.

The final steps of lymphocyte differentiation occur in secondary lymphoid organs where B and T lymphocytes interact with the lymphoid microenvironment. Although numerous studies describe the interactions of murine lymphocytes with dendritic, follicular and other antigen presenting cells, little is known on the interactions between lymphocytes and reticular cells, an important cellular component of spleen stroma. In this work we describe the culturing of complete murine spleen stromas and of two cell lines, Sp-1 and Sp-2, identified as of possible reticular origin, and describe the adhesive interactions between murine lymphocytes and human lymphoid cells with murine spleen stromal cells. FACS analysis indicates that the Sp-1 cell line shows a single cell type expressing VCAM-1 and CD44 constitutively. They do not express any of the markers described for follicular cells, interdigitating cells, macrophages or endothelial cells. Our data suggests that these cells represent a population of spleen reticular cells. The Sp-2 cell line shows two phenotypically different cell types that grow in association. FACS analysis demonstrates that both cell types express VCAM-1 and CD44 constitutively, but that they can be differentiated by the expression of CD11b and FcR. These data suggest that the Sp-2 cell line is composed of one type of stromal cell growing over an adherent layer of reticular cells. Furthermore, analysis of the non-B non-T cell fraction prepared from murine spleen shows that approximately 30% of these cells correspond to the CD44/VCAM-1 double positive cells. Murine B and T cells adhere to the complete stromas and to Sp-1 and Sp-2 cell lines. Activation of B cells with LPS had no effect on binding while binding of T cells to complete stromas increased up to threefold after Con-A treatment. Adhesion of human lymphoblastoid Daudi cells to complete spleen stromas is blocked by an anti-(murine) VCAM-1 antibody but not by an antibody to the (human) integrin alpha 4 subunit, while adhesion to the Sp-1 and Sp-2 stromas is blocked by antibodies against both molecules. Also, adhesion of Ramos cells to Sp-2 stromas is inhibited by antibodies to the integrin alpha 4 subunit and to murine VCAM-1. Antibodies to other adhesion receptors such as the integrin beta 2 subunit, ICAM-1 or CD44 have no effect on human cell binding to these stromas. Our results suggest that we have isolated a fraction of splenic reticular cells and that these cells can be cultured as a distinct cell line. The finding that these cells express CD44 and VCAM-1 constitutively and use some of these molecules for lymphocyte binding suggests that spleen reticular cells may be involved in the regulation of normal lymphocyte traffic through the spleen.

Animals↗
PubMed1997

Adjuvant spleen ultrafiltrate immunotherapy in unresectable advanced non small cell lung cancer.

The purpose of this study was to assess the possible benefit of adjuvant spleen ultrafiltrate immunotherapy in advanced lung cancer patients on chemotherapy. Twenty-six patients with inoperable non-small cell lung carcinoma were eligible to this study and randomised divided into two groups. The two groups received additionally to standard chemotherapy--three times once a month--placebo or spleen ultrafiltrate (Prosplen), respectively, given to the patients for 14 days beginning from the 7th day after each chemotherapy. To evaluate the effects of spleen ultrafiltrate, the time of haematological recovery and profile of peripheral blood lymphocytes and clinically number of days with fever and oncological response were documented. During the observation time patients receiving spleen ultrafiltrate had a higher number of leukocytes (p = 0.01) and higher counts (p = 0.03) and percentage of granulocytes (p < 0.001) including nadir values (p = 0.05) in blood. The positive effect was also seen in natural killer cells (p = 0.005) but not in T cells compartment. This could be of clinical significance because patients receiving spleen ultrafiltrate presented less frequently febrile episodes than patients in the placebo group (75 vs. 127 days with body temperature > 38 degrees C, p = 0.007) and had less frequently highly elevated serum C-reactive-protein levels (p = 0.02). Notably, 2 out of 12 patients receiving in addition to chemotherapy spleen ultrafiltrate and 7 out of 14 lacking this adjuvant treatment showed tumor progression during the treatment. Serum C3 levels were associated with progression of disease in both groups (p = 0.03). Overall, spleen ultrafiltrate receiving patients had lower serum C3 values than placebo receiving patients. Serum IgM levels were rather high in the placebo group (p = 0.033). Spleen ultrafiltrate is thought to benefit patients on palliative chemotherapy in advanced metastatic cancers.

Adjuvants, Immunologic↗
PubMed1979

D-penicillamine: mitogenic effect on mouse, rat and human spleen lymphocytes.

The effect of D-penicillamine (D-Pen) on the proliferation of cultures of normal mouse, rat, and human spleen lymphocytes and peripheral blood lymphocytes was examined. D-Pen in concentrations of 2 x 10(-3) M to 8 x 10(-3) M in serum-free and in serum-containing medium resulted in a highly significant incorporation of 3H-TdR by normal mouse and rat spleen cells. Enhanced incorporation of 3H-TdR by normal human spleen cells only occurred in serum-containing medium. D-Pen in concentrations of 10(-4) M to 10(-3) M in serum-free and serum-containing medium resulted in significant inhibition of 3H-TdR incorporation by normal and mitogen-stimulated mouse and rat spleen cells. Doses of D-Pen greater than 2 x 10(-2) M strongly inhibited 3H-TdR incorporation by both normal and mitogen-stimulated mouse, rat, and human spleen cells and peripheral blood cells. The latter cells were not stimulated or inhibited at lower concentrations of D-Pen. Results from cell depletion and enriching procedures (specific antibody +C' cell killing, employment of athymic, nude spleen cells, adherent and phagocytic cell removal, E rosette cell separation procedures) suggested that target cells in the mouse spleen for D-Pen activation are non-adherent B cells whereas the D-Pen responsive cells in the human spleen probably are T cells.

Animals↗