Lymphocyte Migration Patterns Research Articles

Small bowel transplantation: future prospects

The mesenteric lymph nodes and Peyer's patches of the small bowel present a significant target for immune attack in rejection. In addition they have the potential to proliferate and cause graft-versus-host disease (GVHD). Immune-mediated impairment of mucosal barrier function will allow translocation of bacteria from the gut lumen into the blood stream. This factor, with the development of septicaemia, was probably responsible for the majority of deaths in early clinical cases treated with azathioprine and prednisolone. In the experimental situation GVHD and rejection can be studied as separate entities in transplants between parental strain rats and F1 hybrids. The use of immunohistochemical staining with strain specific monoclonal antibodies has enabled the migration pattern of lymphocytes to be studied. Within 24 h of transplantation donor lymphocytes can be detected in the spleen and mesenteric lymph nodes of the recipient. This cell transfer is clearly effected through the blood stream as the severed lymphatics of the grafted bowel take at least 7-10 days to regenerate and from connections with the recipient lymph system. At a practical level impairment of normal lymphatic drainage may cause problems with the absorption of fats and fat-soluble molecules such as cyclosporin. Denervation of the bowel also presents a problem in the initial phase after transplantation, with hypersecretion from the crypts causing diarrhoea. Since the introduction of cyclosporin a small number of successful human transplants have been reported.(ABSTRACT TRUNCATED AT 250 WORDS)

Lymphocyte migration and localisation patterns in rainbow trout Oncorhynchus mykiss, studied using the tracer sample method

The localisation of 3 H-thymidine labelled lymphocytes after intravenous injection was studied in three experimental designs. Firstly, peripheral blood lymphocytes were separated into sIg + and sIg − and injected back into the same fish. After 24 h, and a passage through the thymus, the distribution of sIg + between the spleen and kidney was approximately the same, but more of the sIg − cells localised in the kidney. Secondly, unseparated lymphocytes were injected into non-histocompatible recipients, and studied over time. By 72 h, 42% of the total cells were in the spleen. Thirdly, organ specific lymphocytes were separated into sIg + and sIg − cells and injected into non-histocompatible recipients. It was found that sIg − cells (from the spleen, kidney and thymus) migrated preferentially to the spleen. The conclusion is drawn that the phenomenon of ecotaxis occurs in fish.

Aberrant Lymphocyte Migration Patterns in Systemic Lupus Erythematosus (MRL/l, MRL/n) Mice are Independent of the Micro-Environment

Mice with systemic lupus erythematosus (SLE) have unusual patterns of lymphocyte traffic characterised by diminished uptake of intravenously injected autoimmune cells into lymph nodes. This study examines the influence of the lymphocyte micro-environment on this aberrant migratory behaviour. To evaluate lymph node lymphocyte-endothelial interactions which can affect lymphocyte distribution without the in vivo influence of liver and spleen, the in vitro high endothelial venule (HEV) binding assay was used. Lymph node HEV binding of autoimmune MRL-lpr/lpr (MRL/l) and MRL(-)+/+ (MRL/n) lymphocytes was increased when compared with CBA/T6 lymphocytes and contrasted with diminished lymph node uptake noted in vivo. This was independent of the lymph node source (MRL/l, MRL/n, CBA/T6) onto which the lymphocytes were overlaid. To examine the influence of the microenvironment on in vivo traffic, 21Cr-labelled lymph node cells from normal CBA/T6 mice were injected into sex-matched MRL/l, MRL/n and CBA/T6 recipients. The distribution of cells was the same in each recipient strain suggesting that the micro-environment had little influence on the lymphocyte trafficking profiles of autoimmune mice. This study supports the view that aberrant lymphocyte migration in autoimmune mice results from defects intrinsic to the lymphocyte population and not the micro-environment.

Lymphocyte migration patterns in organ allograft recipients.

A central tenet of immunology is the observation, made 30 years ago, that lymphocytes recirculate continuously between peripheral blood and lymphoid tissues. In recent years, the subject of lymphocyte migration, both under physiological conditions and in states of alloresponsiveness, has become more enigmatic. It lies outside most current topics of immunological investigations, labelling and tracing techniques are problematic, and many experimental findings are phenomenological and difficult to interpret. Indeed, our overall knowledge of the functional differences between the various host lymphoid compartments and their constituent cell populations remains rudimentary. However, as understanding increases regarding the host immunological events responding to an antigenic stimulus such as a graft, with growing definition of the distinctive and interconnecting roles of lymphocyte subpopulations and their products acting on each other to produce graft destruction, the conceptual importance of lymphocyte migration again is becoming obvious. This role includes many facets of immunity such as the effects of antigen specificity, immunologic memory, differential behavior of recirculating or sessile populations, and local and systemic contact between antigen and effector cells. It has become evident that lymphocytes migrate in a non-random and highly dynamic fashion determined by a range of specific and non-specific factors; in the setting of organ transplantation, patterns are profoundly affected by the interrelated cellular and humoral components of the immunological cascade which may lead either to graft rejection or to its prolongation in untreated and immunologically modified recipients, respectively. Thus, the traffic of lymphocytes throughout host lymphoid and non-lymphoid compartments and their activity within these compartments should be considered an integral part of the host immunomodulation triggered by transplantation of histoincompatible tissue. Gradual filling of the gaps in our current knowledge on the mechanistic aspects of this phenomenon will not only contribute to basic science itself, but also should lead to the development of innovative therapeutic approaches to treat graft rejection.

Lymphocyte Migration Patterns in Autoimmune MRL-lpr/lpr Mice: Relationship to Age, Disease Manifestations and Lymphocyte Homing Receptor Expression

We have previously reported that lymphoid cells from systemic lupus erythematosus (SLE) mice with established disease migrate aberrantly. This study evaluates the abnormal lymphocyte migration patterns found in MRL-lpr/lpr (MRL/l) mice in relation to age, disease manifestations and the expression of lymphocyte homing receptors. 51chromium-labelled lymph node cells from MRL/l and from normal histocompatible CBA mice of different ages were injected i.v. into age and sex-matched CBA recipients. Diminished lymph node and increased hepatic uptake of MRL/l compared to CBA cells was evident as early as 6 weeks of age. Abnormalities in lymphocyte migration antedated the appearance of elevated antihistone antibody (AHA) levels but not the development of lymphadenopathy. Using the monoclonal antibody MEL-14, no differences in the expression of lymphocyte homing receptors between MRL/l and CBA lymph node cells were found at any age. Thus abnormalities in lymphocyte migration in MRL/l mice appear as early as six weeks and are not related to changes in homing receptor expression.

Patterns of disease in localized extranodal lymphomas.

The site of origin of lymphoid tissue is an important determinant of lymphocyte migration patterns. The association of gastrointestinal (GI) and Waldeyer's ring lymphoma and the unique lymphocyte migration pattern of gut-associated lymphoid tissue (GALT) have been previously described. To establish whether predictive clinical patterns of disease occur in localized Non-Hodgkin's lymphoma, survival and relapse patterns for 496 patients with stage I and II non-Hodgkin's lymphoma (NHL) treated with loco-regional irradiation (XRT) alone were examined. We identified 139 patients with GALT lymphoma (defined as arising from primitive gut and including Waldeyers' ring, thyroid, and GI lymphomas) and 87 patients with extranodal non-gut-associated lymphoma (ENL). Survival and relapse data were assessed in multifactorial analysis to correct for previously identified other prognostic variables. GALT lymphomas (GALT-L) have a survival advantage compared with other ENL (P = .017) independent of stage and histology. A difference in distant relapse (DR) rate between GALT-L and other ENL (P = .0002) was also identified. The presentation site of localized extranodal NHL is predictive of clinical behavior and is an independent determinant of outcome. This may be an expression of lymphocytic origin and determinants of migration patterns.

Lymphocyte Traffic and the Skin

Lymphocyte migration patterns into and within lymphoid organs and nonlymphoid organs such as the skin as highly regulated and controlled by a variety of factors, some of which we are beginning to understand at a basic cellular and molecular level. In the case of lymphocyte migration into lymphoid organs, recirculatory patterns and preferences appear to be mediated by selective recognition via lymphocyte membrane receptors of distinctive determinants expressed by specialized endothelial cells in different organ sites. A similar model in which endothelial cells in the skin share common determinants with endothelial cells in peripheral lymph nodes can be developed to explain the migration patterns of lymphocytes from the blood into the skin. Cells that have undergone blast transformation in the microenvironment of the skin or peripheral lymph nodes, or both, appear to recirculate selectively within this particular sphere of immunologic influence, whereas other cells that undergo this transformation in the microenvironment of the gastrointestinal tract and its draining mesenteric lymph nodes selectively recirculate within that sphere of immunologic influence. This preferential recirculation increases the probability that the immunologic defense needs of a given region or tissue such as the skin will be met efficiently and appropriately. Finally, some of the phenotypic characteristics favoring the movement of lymphocytes from the bloodstream into the dermis (such as recent stimulation and the presence of T-cell surface markers) also seem to favor lymphocyte migration into or retention in the epidermis. However, under appropriate circumstances, epidermotropism can be exhibited by both major subsets of T cells as well as a variety of non-T cells.

Lung allograft rejection in the rat. II. Specific immunological properties of lung grafts.

The immunological mechanism of lung allograft rejection was studied in inbred rats, in order to explain the rapid progress of the rejection response against RT1-incompatible lung grafts. Histological appearances of the graft and of the recipient's spleen were studied, migration patterns of graft and recipient lymphocytes were assessed, and titers of circulating alloantibodies were determined. Histologically, we discriminated four phases of the rejection response in lung grafts: sequentially the latent, vascular, alveolar, and destruction phases. Early in the vascular phase, recipient lymphocytes primarily infiltrated the bronchus-associated lymphoid tissue (BALT) of the graft, causing a local immune response. Concurrent with these local rejection phenomena in the graft, a strong systemic immune response developed in the recipient's spleen, presumably induced by the great number of lymphocytes that migrated from the graft's BALT into the recipient's lymphoid tissues. We conclude that BALT facilitates a fast and intensive interaction between lung graft and recipient that is likely to accelerate the induction of the rejection response both locally in the graft and systemically in the recipient's lymphoid organs.

The importance of lymphocyte migration patterns in experimental organ transplantation.

Revue. Variations du pool des lymphocytes recirculants et leurs manipulations lors de transplantations experimentales. Applications possibles chez l'homme. Detection precoce du rejet au moyen de leucocytes marques lors de transplantations de rein ou de cœur. Tentatives d'induction de tolerance durable

Distribution of radiolabelled lymph cells in lymph nodes and the migratory properties of blood lymphocytes in sheep.

The results found in this study supported the concept of lymphocyte populations with preferential migratory pathways. Preferential localization of lymphocytes in lymph nodes could not explain the non-random lymphocyte migration patterns observed in sheep. The migration of lymphocytes isolated from the blood was similar to that of lymphocytes in the efferent lymph of a subcutaneous lymph node, but was different from the migration of lymphocytes isolated from efferent intestinal lymph. The subcutaneous lymph node did not need to be selective for lymphocytes which entered from blood. However, there must be a selective entry of a population of lymphocytes with preference for migrating through mesenteric lymph nodes into efferent intestinal lymph in order to observe the differential lymphocyte migration patterns exhibited by the lymphocytes in the efferent intestinal lymph of sheep.

Assessment of in vivo natural antitumor resistance and lymphocyte. Migration in mice: comparison of 125I-iododeoxyuridine with 111indium-oxine and 51chromium as cell labels.

Clearance of IV-injected tumor cells has been correlated with levels of natural killer (NK) cell activity in recipient animals. Studies of in vivo tumor cell clearance strongly suggest a relationship between levels of NK cell activity and antitumor or antimetastatic effector function. This study outlines the applicability of three radiolabels, [125I]iododeoxyuridine, ( [125I]dUrd), indium-111-oxine chelate ( [111In]Ox), and chromium-51 (51Cr), to studies of tumor cell clearance in vivo. The suitability of these labels for analysis of the in vivo migration patterns of normal lymphocytes or thymus-derived T cells cultivated in vitro (CTC) is also discussed. The results indicate that [111In]Ox and 51Cr compare favorably with the more widely used [125]dUrd as radiolabels for the assessment of IV-injected tumor cell clearance from the lungs of mice. The rates of clearance of both [111In]Ox and 51Cr, like that for [125I]dUrd, correlate closely with levels of NK-cell activity of the host. Further studies with [111In]Ox reveal that treatment of recipients with anti-asialo GM1 serum, a regimen known to suppress NK-cell activity, demonstrates the appropriate reduction in isotope clearance from the lungs after NK suppression. However, clearance data obtained by monitoring levels of radioactivity in the liver after IV injection must be viewed cautiously, since the same cells labeled with [111In]Ox and [125I]dUrd had a different pattern of clearance from the liver. The same inconsistencies in clearance were observed when [111In]Ox and [125I]dUrd were injected intrafootpad (i.f.p.). Similar effects were observed when [111In]Ox or 51Cr was applied to studies of CTC migration. Levels of [111In]Ox and 51Cr remained high in the liver after IV injection, while [125I]dUrd was rapidly cleared. Normal spleen or thymic lymphocytes exhibited the expected homing to the spleen after labeling with [111In]Ox, indicating a suitability of this label for migration studies, except possibly in the liver. These results with CTC and normal lymphocytes should be considered during the formulation of immunotherapy protocols based on cell migration data, since the choice of radiolabel can result in widely divergent levels of radioactivity accumulated in some organs, and may not provide an accurate representation of the presence of viable, intact, or functional cells.

Modification of a transplantable colon tumor and immune responses in mice fed different sources of protein, fat and carbohydrate

The effects of different sources of dietary protein, fat and carbohydrate on tumor development and on tests relating to cell-mediated immunity were investigated in male BALB c mice after subcutaneous injection of 8 × 10 4 1,2-dimethylhy drazine (DMH)-induced colon tumor (no. 51) cells. Results indicated that mice fed the milk protein source (especially at the low protein level) had smaller tumors, a higher spleen cell proliferative response to stimulation by phytohemagglutinin (PHA), and greater cytotoxic T-cell activity against the tumor cells than those fed the comparable diets containing protein from the other sources. Peripheral blood lymphocytes only from the milk-fed mice, regardless of tumor presence, exhibited a relatively low response to PHA stimulation, thereby suggesting a dietary effect on the migration pattern of PHA-responsive lymphocytes. The level of protein significantly affected both T-cell and natural killer cell cytotoxicity. The tumor-bearing mice fed the diet containing sucrose (table sugar) had a significantly lower spleen cell response to PHA stimulation than those fed the comparable diet containing dextrin. The level or source of fat did not significantly affect any of the parameters tested in this system.

Lymphocyte migration during the development of regional lymph node anergy in experimental tumor growth

The development of lymph node anergy in Wistar rats to growing Walker carcinoma 256 was studied in vitro using the 51Cr-release cytotoxicity assay. Cell-mediated cytotoxicity to the tumor peaked in draining lymph nodes 11 days after tumor transplantation. By 14 days, the regional lymph node had become anergic to the tumor at a time when cell-mediated cytotoxicity was still increasing in the more distal contralateral lymph node. Lymphocyte migration into resting, cytotoxic, and anergic lymph nodes was analyzed to determine if altered cell migration into the regional lymph node was associated with the development of anergy. Lymphocyte migration was found to be enhanced in both cytotoxic and anergic regional lymph nodes of tumor-bearing animals. It is concluded that lymph node anergy in this experimental tumor system is not related to changes in lymphocyte migration patterns; rather, it is the result of alterations in the microevironment of the lymph node which prevents the expression of cytotoxic effector cells.

Restoration of allograft responsiveness in B rats by interleukin 2 and/or adherent cells.

LEW X BN)F1 cardiac allografts survive 1 wk in unmodified LEW recipients (MST +/- SD = 8.3 +/- 1.2 days) but indefinitely (greater than 100 days) in B rats, produced by 750 R sublethal x-radiation 3 to 4 wk after adult thymectomy and reconstitution with syngeneic bone marrow cells from thymectomized thoracic duct-drained donors. Graft survival appears independent of blocking antibodies and is not mediated by suppressor cells or loss of graft immunogenicity. The unresponsive state is eventually reversed by adoptive transfer of 10(8) spleen cells from nonimmune (MST +/- SD = 27.5 +/- 4.7 days) or alloimmune (MST +/- SD = 21.5 +/- 1.9 days) syngeneic animals. In contrast, concomitant administration of 2 X 10(7) thioglycollate-stimulated peritoneal exudate adherent cells plus 10(8) alloimmune syngeneic spleen cells produced acute allograft rejection in 50% of B recipients within 10 to 12 days, while in every instance grafts underwent acute rejection (MST +/- SD = 10.0 +/- 1.4 days) in B recipients treated with interleukin 2 (IL 2) rich conditioned supernatant plus sensitized cells. In vitro studies revealed that adherent cells from B rats possessed less than 50% the capacity of adherent cells from normal animals either to support the Con A-stimulated uptake of 3H-thymidine by splenic T cells or to promote production of IL 2 by spleen cells depleted of adherent cells. Altered lymphocyte migration patterns in B recipients may also contribute to prolonged allograft survival secondary to adherent cell dysfunction.

Migration patterns of lymphocytes in untreated and immunologically manipulated recipients of organ allografts.

We have investigated the migration patterns of normal LEW rat splenic lymphocytes (SLs) radiolabeled in vitro with [2-3H]adenosine and adoptively transferred i.v. into LEW hosts bearing LBNF1 heterotopic cardiac allografts. Twenty-four hours after cell transfer, the animals were killed and the radioactivity of all lymphoid and nonlymphoid tissues measured in a beta counter. The following experimental groups were studied: group 1, untreated recipients acutely rejecting their grafts at 7 days after transplantation; group 2, actively and passively enhanced recipients bearing long-term surviving grafts, at 7, 14 to 18, and 25 days; group 3, cyclosporin A-treated recipients; and group 4, B rats, each bearing indefinitely surviving grafts, at 7 and 20 to 30 days. In group 1, 28% of recoverable activity was found in spleen and 22% in mesenteric and peripheral lymph nodes. In animals with well functioning grafts of groups 2 and 3, SLs accumulated in both organs equally (25 to 27%). In animals in group 4, SLs migrated primarily to lymph nodes (30%) and away from spleen (20%). Sequestration in nonlymphoid tissues of animals experiencing graft rejection was higher than those with prolonged or indefinitely surviving hearts. By using mouse anti-rat monoclonal antibodies, the quantitative relationship between T cell subpopulations in transferred cell suspensions and in lymphoid organs of grafted hosts was also assessed. Lymphocyte migration patterns are influenced dramatically by the immunological status of recipients of vascularized organ allografts.

Mechanisms of immunosuppression in tumor-bearing mice: a multifactorial analysis.

Neoplastic growth results not only from the inherent proliferative potential of the neoplasm, but also from the neoplasm's ability to survive within a host whose immune responses may be activated to destroy it. It has been proposed that several mechanisms enable a neoplasm to outgrow the host. The present work describes a system for probing tumor-associated immunosuppression by utilization of the delayed hypersensitivity response to dinitrochlorobenzene (DNCB). Mice of the Balb/c and C3H/HeJ strains were sensitized with a subcutaneous injection of 2 mg DNCB followed 5-10 days later by the subcutaneous injection of a 0.05 mg challenge dose of DNCB in the hind footpad. The footpad response measured 24 hours after challenge is a delayed hypersensitivity reaction by virtue of the histology and kinetics of that response and because it is transferred by spleen cells but not by serum. DNCB sensitization is inhibited by spontaneous, chemical- and virus-induced syngeneic tumors in Balb/c and C3H/HeJ mice. Three mechanisms of immunosuppression are elucidated in the present experiments: (1) serum from tumor-bearing mice inhibits primary sensitization of normal mice to DNCB upon passive transfer; (2) spleen cells from tumor-bearing mice inhibit primary sensitization of normal mice to DNCB and appear to be macrophages in the C3H/HeJ system; and (3) an alteration of lymphocyte migration patterns exists in tumor-bearing mice that removes effector cells from the blood and prevents the initiation of a footpad response. The interrelationships of these three mechanisms are discussed. Correlations are also made with tumor-associated immunosuppression observed in cancer patients.

Migration patterns of lymphocytes from recipients of organ allografts. I. The unmodified host.

We have investigated in the rat, serial dynamics and homing patterns of different lymphocyte populations from unmodified recipients of heterotopic cardiac allografts and have compared these kinetics to those of lymphocytes from isografted or nongrafted control animals. Sensitized lymphocytes were labeled in vivo and in vitro. Groups of allografted rats were inoculated in vivo with [6-3H]thymidine serially following transplantation. Alternatively, lymphocytes were isolated from all lymphoid compartments of recipient animals at intervals following grafting and labeled with 51Cr. Spleen cell suspensions were further fractionated into nylon-wool adherent (SIg+) and nonadherent (T) populations, which were then differentially labeled with [2-14C]thymidine and [6-3H]thymidine, respectively. All cell groups were then transferred into normal syngeneic animals, and migration patterns into all lymphoid and nonlymphoid organs were assessed 24 hr later. In vivo labeling of sensitized lymphocytes showed a significant increase in radioactivity in recipient lymph nodes, spleen, and grafted heart during the period of rejection at 8 days, compared to isograft or normal controls. Dynamics of in vivo labeled lymphocytes from allografted rats differed markedly from controls; total radioactivity of blood and all lymphoid organs of test rats rose progressively (P < 0.001) until the time of rejection, with reciprocal decrease of accumulated radioactivity in nonlymphoid tissues. Distribution and kinetics of all lymphoid cell groups with the exception of those from bone marrow were comparable following transfer into test animals, invariably increasing in lymphoid tissues during the period of rejection, and decreasing in reciprocal relationship in nonlymphoid tissues. Results from double-labeled nylon-wool nonadherent and adherent cells showed that the T to B cell ratio increased steadily in lymph nodes, Peyer's patches, and spleens of test animals between 2 and 8 days, and declined in blood and small intestine.

Effect of infections with swine fever virus on immune functions I. Response of lymphocytes from blood and lymphoid organs from infected and normal pigs to anti-immunoglobulin serum and protein A

Pigs exposed to a low-virulent strain of swine fever virus (SFV) developed an inapparent infection. At times when a transient leucopenia occurred, the peripheral blood lymphocytes (PBL) were unresponsive to the mitogenic stimulus of anti-immunoglobulin serum (anti-Ig) and protein A. Pigs lethally infected with a virulent SFV showed leucopenia and unresponsiveness of PBL to anti-Ig and protein A from 2 days post infection until death. This suggests a defect in B lymphocyte function in pigs infected with SFV. The unresponsiveness to anti-Ig appeared not to be caused by a reduced ability of lymphocytes to redistribute their receptors into caps, the presence of suppressor cells or absence of surface immunoglobulin bearing lymphocytes in the peripheral blood. A direct action of the virus itself also seemed unlikely. Lymphocytes from spleen reacted as PBL. However, lymph node cells did not lose their capability to respond to anti-Ig. These data suggest that a change in the migration pattern of anti-Ig responsive lymphocytes could account for the observed unresponsiveness of PBL and spleen lymphocytes to anti-Ig.

Migration pattern of splenic lymphocytes after local labelling of the pig spleen with 3H-cytidine.

In normal young pigs splenic lymphocytes were selectively labelled by injecting tritiated cytidine into a splenic artery. 10 h later several lymphoid and non-lymphoid organs were investigated for spleen-derived lymphocytes by autoradiography. The relative and absolute organ distribution of the labelled cells was determined. Labelled lymphocytes appeared rapidly in the peripheral blood reaching a mean labelling index of 4.8%. More splenic lymphocytes were found in thymus dependent areas than in thymus independent areas of lymphoid organs. Nearly 40% of all emigrated lymphocytes homed to lymph nodes and only about 1% in the thymus. A surprisingly high number of splenic lymphocytes were located in the bone marrow and the lung.

Splenic lymphocytopoiesis and migration pattern of splenic lymphocytes

The spleens of young pigs were selectively labeled with tritiated thymidine ([ 3H]-TdR) and the relative and absolute numbers of labeled lymphocytes found 24 hr later in different lymphoid and nonlymphoid organs were determined autoradiographically. It was deduced that about 4.6 × 10 9 lymphocytes (that is, about 15% of all splenic lymphocytes) are produced by the spleen per day and about 17% of the newly formed lymphocytes leave the spleen within the first day of labeling. Spleen-derived lymphocytes could be found in relatively high numbers in the lymph nodes, blood, gut-associated lymphoid tissues, and, surprisingly, in the bone marrow, whereas the concentration in the thymus was very low. In a second series, pigs were labeled with [ 3H]TdR and only the spleen was excluded from labeling. The labeling index of splenic small lymphocytes was about 10% 1 day later, indicating a high rate of influx of newly formed lymphocytes into the pig spleen. The spleen of the young pig is an important lymphocytopoietic organ and exports and imports newly formed lymphocytes at high rates.