Lymphocyte migration in internally irradiated animals. Effect of internal beta radiation on the migration of spleen lymphocytes in CBA mice

The syngeneic transfer system was used to study migration of 51Cr-labelled spleen lymphocytes in mice after incorporation of beta-emitter, 35S-methionine. Migration of 51Cr-labelled lymphocytes to lymph nodes was stably decreased, and to liver, kidneys and lungs increased. The lymphocyte migration impairment was associated with the influence of beta-radiation on both the migratory properties of cells and the factors of their microenvironment responsible for the lymphocyte migration within the mouse body. No distinctions were observed in the character and manifestation of disturbances of the lymphocyte migration after the injection of 35S-methionine and gamma-emitter, 75Se-selenomethionine.     

Lymphocyte migration in internally irradiated animals. Effect of internal gamma radiation on the migration properties of spleen lymphocytes labeled with 51Cr

In experiments on CBA mice it was shown that migration of 51Cr-labeled spleen lymphocytes, injected intravenously, to lymph nodes of intact recipients was suppressed 6-24 months after the administration of a radiopharmaceutic preparation of selenium-75-selenomethionine in a quantity forming the doses of 1 Gy and 1.5 Gy absorbed within the whole body and lymphoid organs, respectively. Migration of labeled lymphocytes to the liver, kidneys and lungs, as well as their retention in the circulating blood, were increased. As the result of the migration disorders the specific affinity of lymphocytes for peripheral lymphoid tissue decreased.     

In vitro studies on transplantation immunity. I. MIF production by sensitive lymphocytes in mice

Sensitized lymphocytes from mice immunized with skin homografts produce migration inhibitory factor upon incubation with lymphocytes (antigen) from the sensitizing strain. The MIF is produced within 14 hr following incubation of sensitized lymphocytes and antigen. In this reaction, antigenic specificity is a prerequisite for MIF production; however, the action of MIF transcends the strain barrier. Also, MIF produced in homograft reactions in mice inhibited the migration of peritoneal cells from normal guinea pigs. Finally, lymphocytes from mice bearing skin homografts do not develop the capacity to produce MIF prior to the rejection of the sensitizing skin grafts.     

Virus-induced alterations of lymphoid tissue. IV. The effect of Newcastle disease virus on the fate of transfused thoracic duct lymphocytes

⁵¹Cr-labeled thoracic duct lymphocytes were briefly incubated at 4 °C with Newcastle disease virus (NDV) and then infused into syngeneic rats. Virus diverted the homing of many donor cells from lymph nodes and spleen to the liver. Evidence was obtained suggesting that some NDV-treated lymphocytes initially trapped in the liver subsequently migrated into the lymph nodes. The results imply that NDV transiently interrupts the normal route of lymphocyte migration. Alterations in lymphocyte distribution were mediated by attachment of virus to the cell surface and were the same as those induced by incubating lymphocytes with V. cholera neuraminidase before infusion. It appears that reactions involving 2–3′ and/or2–8′ linked sialyl residues on the surface of recirculating lymphocytes can markedly affect their distribution in the body.     

Kinetics of indium-III labelled lymphocytes in normal subjects and patients with Hodgkin's disease.

The distribution in the body and the circulation in the blood of autologous lymphocytes labelled with indium-III were studied in two normal subjects and two patients with Hodgkin''s disease. Four hours after injection radioactivity was identified in the spleen, liver, and bone marrow. Radioactivity, followed by imaging and whole body scanning, began to appear in the lymph nodes four to 18 hours after injection, and some, though not all, lymph node groups in the body could be readily visualised. There were no differences between the normal subjects and the patients with Hodgkin''s disease. The pattern of clearance of radioactivity from the blood was consistent with a normal circulation between blood and lymphoid tissues of the labelled lymphocytes. Since indium-111 stays firmly attached to the cell, it seems an ideal label for studying lymphocyte kinetics, and the use of this technique may have further clinical application.     

Direct leukocyte migration inhibition by myelin basic protein in exacerbations of multiple sclerosis.

Studies in 13 normal subjects, 9 patients with multiple sclerosis (MS) within 3 weeks of exacerbation and 16 others 1 to 6 months after onset were carried out for evidence of cell-mediated hypersensitivity to myelin basic protein. Ten patients with stroke and 10 with Guillain-Barré syndrome were studied as additional controls. Peripheral leukocytes obtained by leukapheresis were packed into capillary tubes and allowed to migrate out onto glass in the presence or absence of myelin basic protein. Cells of patients within 3 weeks of an MS episode gave a mean migration index of 68 +/- 9%, and those 1 to 6 months after onset, 93 +/- 21%. For the entire MS group the mean index was 88 +/- 20%, for those with Guillain-Barré, 103 +/- 7%; and for the stroke patients, 107 +/- 11%. Results for the acutely ill MS patients were significant (P less than 0.005). The data are similar to those obtained using the migration inhibition factor assay but show that sensitized lymphocytes also elaborate a second mediator during acute exacerbations of illness. These observations strengthen evidence that sensitization to this potent encephalitogen occurs simultaneously with exacerbations of clinical illness.     

Suppressor cells in homograft tolerant rats.

If sufficient normal syngeneic lymphocytes to effect skin graft rejection are transferred to homograft tolerant rats, a prolonged period elapses before lymphoid cells from the recipient acquire normal levels of GvH responsiveness against tissues of which the donor was previously tolerant (Silvers and Billingham, 1970; Elkins, 1972; Miyamoto and McCullagh, 1974). Although the ability of lymphoid populations of such animals to mount GvH reactions can be demonstrated to reside in donor type cells during the weeks immediately after transfer, reactive cells are ultimately derived from the host itself (Elkins, 1973; Miyamoto and McCullagh, 1974). Not only are lymphoid cells from tolerant rats which have been injected recently with normal lymphocytes poorly responsive in a GvH assay, but they have been observed in some experiments to suppress the GvH activity of normal syngeneic lymphoid cells (Elkins, 1972; Atkins and Ford, 1972). It is not clear whether the cells mediating suppression of the normal lymphocytes were derived from the tolerant host itself or, alternatively, from the normal lymphocytes injected into it to terminate the tolerant state. The present experiments sought to delineate the origin of any suppressor cells within populations of lymphocytes collected from rats in which tolerance had recently been terminated. The indicate that suppression of the normal donor cells within such populations may be exerted by cells derived from the tolerant host.     

Lymphocyte recruitment in delayed-type hypersensitivity. The role of IFN-gamma

Lymphocytes are recruited out of the blood into delayed-type hypersensitivity (DTH) reactions, but the factors controlling their migration are poorly understood. Our previous studies have shown that IFN-alpha/beta, its inducers, and T cell lymphokines can induce lymphocyte migration into the skin after intradermal injection. The present studies were designed to determine the effect of rIFN-gamma, IL-1, and anti-IFN-gamma on lymphocyte recruitment into DTH. Small peritoneal exudate lymphocytes, which preferentially migrate to inflammatory sites, were labelled with 111In and injected i.v. into rats. The intradermal injection of IFN-gamma stimulated the migration of these lymphocytes into the skin. IL-1 induced very little migration by itself, but enhanced the effect of IFN-gamma. Kinetic analysis demonstrated that the migration of lymphocytes to IFN-gamma was rapid, with a peak at 6 h, whereas migration into a DTH reaction was minimal for the first 8 h and reached a peak 24 h after intradermal injection. Polyclonal rabbit anti-IFN-gamma anti-serum, and a Mab to IFN-gamma, DB-2, could almost completely block lymphocyte migration induced by IFN-gamma. Furthermore, DB-2 inhibited lymphocyte recruitment into DTH reactions by 50 to 90%. This Mab did not affect migration in response to IFN-alpha/beta, although it partially inhibited the response to polyI:C. The effect of IFN-gamma on lymphocyte recruitment was not specific for small peritoneal exudate lymphocytes, because both spleen T cells and lymph node cells migrated in response to IFN-gamma and DB-2 inhibited the recruitment of splenic T cells to DTH. Thus, IFN-gamma is a potent stimulator of lymphocyte migration into the skin and a major mediator of lymphocyte recruitment into DTH.     

In vitro studies on transplantation immunity. II. The migration inhibition in homograft reactions in guinea pigs

The migration of peritoneal exudate cells from guinea pigs exhibiting transplantation immunity is inhibited in the presence of donor antigens. This inhibition of migration is demonstrable whether the donor transplantation antigens are presented in the form of viable cells (peritoneal exudate cells) or as particulate subcellular antigens (spleen microsomes). A greater degree of inhibition was observed when transplantation immunity was induced with lymphoid cells in Freud's adjuvant compared to sensitization with orthotopic skin grafts. There was no inhibition of migration in mixtures of normal allogeneic cells or when peritoneal cells from guinea pigs exhibiting tuberculin hypersensitivity were mixed with similar cells from normal animals. Finally, supernatants from cultures of sensitive lymphocytes plus donor antigens inhibited the migration of normal peritoneal cells indicating the presence of migration inhibitory factor (MIF) activity.     

Site-selective homing of antigen-primed lymphocyte populations can play a crucial role in the efferent limb of cell-mediated immune responses in vivo

Lymphoid cells of the mammalian immune system exhibit special migratory properties within their in vivo environment. This fundamental characteristic is important to the protection of the organism from infection and neoplastic transformation by a process termed immunologic surveillance. The importance of lymphocyte recirculation was addressed by investigating the role of site-selective homing of lymphocytes during the efferent phase of an in vivo adoptively transferred immune response. We approached this problem by using pertussis toxin (PT), a known inhibitor of lymphoid cell migration in vivo. PT is a protein secreted by the bacterium Bordetella pertussis, which induces a selective and long-lasting inhibition of the emigration of lymphocytes from the bloodstream into solid tissue. In this study, we demonstrate that the blockade of lymphocyte extravasation potential mediates inhibition of certain cell-mediated immune responses in vivo. We investigated the effect of PT on the extravasation of antigen-primed lymphocytes into solid tissue. The results show that the loss of lymphocyte homing potential after in vitro treatment of the primed cells with PT is accompanied by an inhibition of antigen-specific contact hypersensitivity after adoptive transfer. This result suggests that the process of lymphocyte extravasation into nonlymphoid tissue sites such as the skin shares fundamental similarities to the selective localization of circulating lymphocytes to lymph nodes. Furthermore, the inhibition of contact hypersensitivity observed after the i.v. adoptive transfer of PT-treated antigen-primed cells could be circumvented by transferring the PT-treated cells directly into a tissue site with the relevant antigen. In contrast, no inhibition in the number of antibody-forming cells present within the spleen was observed after an adoptive transfer of PT-treated sheep red blood cell-primed lymphocytes, a result that is in agreement with radiotracer data. Thus, the inhibitory effect of PT on the adoptive transfer of contact hypersensitivity was established to be a direct result of the PT-mediated alteration of cellular migration, because PT inhibits the entrance of lymphocytes into specific tissue sites without inhibiting other cellular functions. This conclusion is additionally supported by experiments that showed that lymphocytes that have been pretreated with PT exhibit normal in vitro responses, as assayed by mitogenesis in response to concanavalin A and by the differentiation and function of alloantigen stimulated or hapten-specific cytotoxic T lymphocytes.(ABSTRACT TRUNCATED AT 400 WORDS)     

5-氮胞苷对贵州小型猪淋巴细胞DNA损伤及修复的影响

目的　研究贵州小型猪淋巴细胞对化学物或药物引起的DNA损伤及修复影响的反应。方法　用单细胞凝胶电泳技术检测比较 5 氮胞苷对PHA刺激和未刺激淋巴细胞的DNA损伤及其修复过程。结果　 5 氮胞苷引起未刺激淋巴细胞明显的DNA泳动 (彗星尾 ) ,经修复孵育 2h后 ,DNA泳动与孵育前比较无显著差异 ,而 5 氮胞苷引起的刺激细胞DNA泳动经 2h修复孵育后与孵育前比较显著减少。结论　 5 氮胞苷引起贵州小型猪未刺激淋巴细胞DNA损伤经 2h孵育未能修复 ,而刺激细胞的DNA损伤明显修复。     

T and B lymphocyte migration into syngeneic tumors.

The migration of splenic T and B lymphocytes into syngeneic tumors undergoing immunologic rejection was investigates. Spleen cells were obtained from normal BALC/c mice or BALB/c mice bearing tumors induced by murine sarcoma virus (MSV). Either whole spleen cells or immunoabsorbent purified T and B cells were radiolabeled with sodium chromate-51 and injected i.v. into normal or MSV inducted-tumor bearing syngeneic recipients. Twenty-four hours later the recipient mice were sacrificed and radioactivity was assessed for tumor, contralateral normal muscle, the lymph nodes draining the tumor and contralateral draining lymph nodes, peripheral lymph nodes, spleen, and liver. Both T and B lymphocytes from either normal or MSV tumor-bearing animals show greatly increased migration into the tumor when compared with normal muscle. Migration of T cells from both normal and MSV tumor bearers was 30 times that of migration to normal muscle. B cells from tumor-bearing mice, on the other hand, localized in the tumor itself only 50% as frequently as did B cells from normal animals. In addition, T cells from MSV tumor bearers were found in the highest proportion in the lymph node draining the tumor site. We conclude that T and B lymphocytes from either normal or tumor-bearing mice migrate to a syngeneic tumor undergoing immunologic rejection. In contrast, the migration of both T and B cells from tumor-bearing animals was decreased to the peripheral lymph nodes at the time of maximum tumor growth.     

Cellular immunity in man: correlation of leukocyte migration inhibition factor formation and delayed hypersensitivity

The formation of leukocyte migration inhibition factor (MIF) by the lymphocytes of 13 normal persons immune to the protein antigen keyhole limpet hemocyanin (KLH) has been investigated. KLH-induced MIF formation expressed as percent migration was compared with delayed hypersensitivity, antibody, and in vitro lymphocyte blastogenic responses to this antigen. Individuals were studied 404–840 days (median 540 days) after their last exposure to KLH. Nine persons had delayed hypersensitivity to KLH and 10 had circulating KLH antibody. The lymphocytes of 11 showed an in vitro blastogenic response to KLH stimulation, while the lymphocytes of nine produced MIF after KLH stimulation. The mean percent migration for the subjects with KLH delayed hypersensitivity was 48.2 (range 20.4–70.4) compared with 133 (range 120–161) for the four persons who did not have KLH delayed hypersensitivity (P < 0.05). The correlation coefficient between the precent migration and delayed hypersensitivity was ?0.78 (P < 0.01). No correlation was demonstrated between migration inhibition and the other parameters of immunity.     

Chemotactic activity of pertussis toxin on murine lymphocytes. Its potential role on lymphocyte accumulation in the lung

The effects of pertussis toxin on lymphocyte migration were studied in vitro. In this study pertussis toxin significantly stimulated lymphocyte migration at concentrations of 0.1 and 1 microgram ml-1 using a microchamber and the leading-front method. Checkerboard analysis demonstrated that pertussis toxin causes directed migration of lymphocytes (chemotaxis). Heat-treatment pertussis toxin abolished its capacity to cause this migration. When murine lymphocytes were preincubated with different concentrations of pertussis toxin, an inhibition of chemotaxis at the dosages of 0.1 and 1 microgram ml-1 was observed. On the other hand, lymphocytes derived from mice treated with pertussis toxin were not inhibited after subsequent exposure to pertussis toxin in vitro. Since lymphocyte accumulation in the lungs of mice treated with pertussis toxin has been well demonstrated, the results of our study could suggest a chemotactic activity of pertussis toxin in determining accumulation of lymphocytes in this organ.     

Cell-mediated immunity in interstitial nephritis. II. T lymphocyte effector mechanisms in nephritic guinea pigs: analysis of the renotropic migration and cytotoxic response.

The present studies investigate the effector role of lymphocytes in guinea pigs with an interstitial nephritis. Several observations were made relative to a number of functions expressed by these cells. The results of adoptive cell migration studies suggest that a subpopulation of T cells in nephritic animals traffic renotropically to either normal or damaged kidneys on transfer. Similar lymphocytes were also tested in vitro for direct effector function by utilizing target kidney cell monolayers in a cell-mediated cytotoxicity assay. The kinetics of the observed cytotoxic response were studied over the life span of nephritic animals. Optimal target-cell lysis occurred 12 to 17 days after sensitization, simultaneous with the onset of active histopathologic changes. The cytotoxicity was stoichiometrically titratable and relatively specific for fetal kidney tissue. In addition, cells from the spleen or lymph nodes of diseased animals effectively suppressed this cytotoxic response. These findings demonstrate that a diverse population of T lymphocytes are both capable of damaging the renal interstitium as well as modulating effector-cell functions on a regional basis with the immune system.     

Chemotactic activity of pertussis toxin on murine lymphocytes. Its potential role on lymphocyte accumulation in the lung

Abstract The effects of pertussis toxin on lymphocyte migration were studied in vitro. In this study pertussis toxin significantly stimulated lymphocyte migration at concentrations of 0.1 and 1 μg ml⁻¹ using a microchamber and the leading-front method. Checkerboard analysis demonstrated that pertussis toxin causes directed migration of lymphocytes (chemotaxis). Heat-treatment of pertussis toxin abolished its capacity to cause this migration. When murine lymphocytes were preincubated with different concentrations of pertussis toxin, an inhibition of chemotaxis at the dosages of 0.1 and 1 μg ml⁻¹ was observed. On the other hand, lymphocytes derived from mice treated with pertussis toxin were not inhibited after subsequent exposure to pertussis toxin in vitro. Since lymphocyte accumulation in the lungs of mice treated with pertussis toxin has been well domenstrated, the results of our study could suggest a chemotactic activity of pertussis toxin in determining accumulation of lymphocytes in this organ.     

Electron microscopic study of Ag-protein localization in the nucleoli of human lymphocytes

The present study was undertaken to provide more information on the ultrastructural localization of a silver reaction in normal resting and stimulated lymphocytes as well as leukaemic resting lymphocytes. The results obtained indicated that in the ring-shaped nucleoli of normal mature lymphocytes silver stained proteins (SSPs) were present mostly within single fibrillar centers. In the nucleoli of lymphocyte cultures, being in the presence of phytohemagglutinin (PHA) for 6--72 hours, SSPs formed finger or loop-like protrusions from fibrillar centers towards the adjacent areas of the nucleoli. In the ring-shaped nucleoli of mature leukaemic lymphocytes SSPs are present not only within fibrillar centers, but also in protrusions diverging from fibrillar centers into the surrounding peripheral nucleolar ring. In this respect the nucleoli of leukaemic mature lymphocytes were similar to normal lymphocytes shortly after mitogen stimulation.     

Lymphocytes bearing Fc receptors for IgE. II. Induction of Fcepsilon-receptor bearing rat lymphocytes by IgE.

The proportion of lymphocytes bearing receptors for IgE (FcepsilonR) markedly increased after infection of rats with Nippostrongylus brasiliensis (Nb). The FcepsilonR-bearing lymphocytes from the infected animals bound more IgE-coated erythrocytes in rosette assay than FcepsilonR-bearing cells from normal rats, suggesting that the number of FcepsilonR per cell may also increase following the infection. In contrast, the number of IgE-receptors on peritoneal mast cells did not change after Nb infection. The increase in the proportion of FcepsilonR-bearing lymphocytes in Nb-infected rats is probably due to an increased concentration of IgE in the environment. The proportion of FcepsilonR-bearing cells in normal rat lymphocyte suspensions increased by culture of the cells with rat IgE of 1 microgram/ml or higher concentration. Other immunoglobulins such as rat IgG, human IgE, or rabbit IgG failed to induce either FcepsilonR-bearing cells or FcgammaR-bearing cells. It was also found that induction of Fc receptors by rat IgE is confined to FcepsilonR. Kinetic studies on the induction of FcepsilonR-bearing lymphocytes in vitro showed that the proportion of these cells in lymphocyte suspensions increased within 8 hr incubation with rat IgE but not within 4 hr. Evidence was obtained that both RNA synthesis and protein synthesis, but no DNA synthesis, are required for the induction of FcepsilonR-bearing cells or the expression of the receptors on the cell surface.     

Endothelium-lymphocyte interactions in vitro. II. Adherence of allergized lymphocytes.

Peripheral blood lymphocytes from skin graft-sensitized pigs will adhere in vitro to fresh donor-type large vessel endothelium, but do not spread out or migrate. Similar cells will however spread out on and migrate through monolayers of cultured donor-type aortic endothelium to a significantly greater extent than nonallergized lymphocytes. Cells sensitized in mixed lymphocyte culture at first exhibit a nonspecific increase in adherence and migration correlated with increased thymidine uptake, but after more prolonged incubation adherence becomes specific for stimulator-type endothelium. It is suggested that lymphocyte infiltration of an allograft in the presence of circulating sensitized cells involves a combination of nonspecific lymphocyte adhesion to endothelium, antigenic stimulation of “primed” cells to increased motility, endothelial penetration and lymphokine production, and soluble-factor-mediated stimulation of migration by nonsensitized cells.     

Changes in the electric charge of blood lymphocyte populations after secondary immunization with tetanus antitoxin in man]

The electrophoretic mobility of circulating lymphocytes has been studied in normal human subjects after immunization by tetanus toxoid. The mean migration speed was shown to increase, particularly two and three days after secondary immunization. This increase appeared to be due to the elevation of percentage of T cells migrating at 1.20 and 1.35 micrometer. sec.-1v.-1 cm. (active rosettes-forming cells), with a decrease of the percentage of B cells and T lymphocytes migrating at 1.10 micrometer. sec.-1v.-1 cm. The return to the anterior status was observed between day 4 and 8 after immunization.