Locomotor responses of human CD45 lymphocyte subsets: preferential locomotion of CD45RO+ lymphocytes in response to attractants and mitogens.

The CD45RO+ population of lymphocytes from human blood contains a higher proportion of locomotor cells than the CD45RA+ population. Direct from blood there were few locomotor lymphocytes (< 15%), but, among these, a higher proportion of CD45RO+ than of CD45RA+ cells responded to the chemotactic stimuli, foetal calf serum (FCS) and interleukin-2 (IL-2) in polarization assays. Likewise, after overnight culture, a higher proportion of CD45RO+ cells responded to IL-8. Culture for 24-72 hr in activators such as anti-CD3, purified protein derivative (PPD), phytohaemagglutinin (PHA), concanavalin A (Con A), pokeweed mitogen (PWM) or in an allogeneic mixed leucocyte reaction (AMLR) increased the proportion of locomotor lymphocytes to 20-60%, and the CD45RO+ subset showed proportionately more polarized cells than the CD45RA+ subset after culture with all the above activators. Preferential migration of CD45RO+ cells into collagen gels was also seen after culture in antigenic stimuli (PPD or AMLR) but not with polyclonal activators (alpha CD3 or Con A). Double labelling showed that, within the CD4+ and CD8+ subsets, antigen-stimulated CD45RO+ T cells invaded collagen gels in higher proportions than CD45RA+ T cells. Clustering of lymphocytes with accessory cells is an essential prerequisite for locomotion and, after culture in alpha CD3, CD45RO+ lymphocytes were found preferentially in clusters with monocytes. In all of the above populations, CD45RO+ lymphocytes were larger in size. These findings suggest that, not only selective adhesion to vascular endothelium as reported earlier, but also selective locomotion recruits CD45RO+ lymphocytes into sites of inflammation.     

Enhanced adhesion of autologous lymphocytes to gamma-interferon-treated human endothelial cells in vitro

An adhesion assay was developed using human umbilical vein endothelial cell cultures and autologous and allogeneic lymphocytes separated from cord blood. Endothelial monolayers cultured in plain or gamma-interferon (IFN gamma)-supplemented medium were cocultured with lymphocytes for 1 h and non-adherent lymphocytes removed by washing. Autologous and allogeneic lymphocytes exhibited significantly increased adhesion with IFN gamma-treated cells compared with untreated controls. The increased adhesion to IFN gamma-treated endothelial cells was significantly inhibited when autologous or allogeneic lymphocytes were pre-treated with saturating amounts of an anti-CD4 monoclonal antibody. The results indicate that IFN gamma enhances lymphocyte binding to endothelium and that the CD4 molecule may be involved in this process. This could be an important mechanism in targetting the migrating of T-helper (Th) cells to areas of chronic inflammation and in antigen presentation by endothelial cells.     

Interactions between interleukin-2-activated lymphocytes and vascular endothelium: binding to and migration across specialized and non-specialized endothelia.

A prerequisite for the successful immunotherapy of solid tumours with interleukin-2 (IL-2)-activated lymphocytes is their ability to home to the tumour tissue. Lymphocyte homing is a complex process which is known to involve at least two independently regulated events: adhesion to the luminal surface of vascular endothelium and the subsequent transendothelial migration of lymphocytes. In this study we have used an in vitro model of lymphocyte homing which employs specialized high endothelium to ask whether IL-2-activated lymphocytes are able to migrate across vascular endothelium in order to leave the blood vessel. Both the adhesion of IL-2-activated cells and their migration across monolayers of cultured high endothelial cells (HEC) were increased in comparison with non-activated lymphocytes. The adhesion of IL-2-activated lymphocytes was mediated by lymphocyte function-associated antigen-1 (LFA-1) and a very late activation antigen-4 (VLA-4)-related pathway. LFA-1-dependent adhesion was mediated by ligands on HEC other than the intercellular adhesion molecule-1 (ICAM-1) and the VLA-4-related pathway was mediated by ligands other than the CS1 domain of fibronectin. HEC-adherent lymphocytes were enriched in natural killer (NK) cells and CD8+ T cells which are known to be the tumour-cytotoxic cells in IL-2-activated lymphocytes. However, there was no evidence of cytotoxicity towards the endothelial layer using a syngeneic model. The interaction of IL-2-activated lymphocytes and endothelial cells was not specific for high endothelium since equal numbers of activated lymphocytes bound to and migrated across aortic endothelium. The inability of IL-2-activated lymphocytes to discriminate between high endothelium and non-specialized 'flat' endothelium could be responsible for the widespread dissemination of the cells throughout the body following their adoptive transfer and the unwanted side-effects at non-involved sites.     

Surface immunoglobulin-positive T lymphocytes in HIV-1 infection: relationship to CD4+ lymphocyte depletion

T lymphocytes bound to autologous immunoglobulin (surface Ig + T cells) and serum antibodies that bind to allogeneic lymphocytes have been detected in HIV-1-infected individuals, but their significance in the immunopathogenesis of HIV-1 infection is uncertain. We tested peripheral blood from HIV-1-infected individuals to determine if surface Ig+ T cells are specific for HIV-1 infection and are associated with CD4+ lymphocyte depletion. The majority of HIV-1-infected individuals contained substantial numbers of circulating surface Ig+ T cells. The presence of such cells was restricted to seropositive individuals and not related to risk factors associated with the acquisition of HIV-1 infection. Autologous immunoglobulin was detected on both CD4+ and CD8+ cells in all patients tested. Most individuals with surface Ig+ T lymphocytes also had serum anti-T-lymphocyte antibodies. The presence of surface Ig+ T lymphocytes correlated significantly with lower absolute CD4+ lymphocyte counts only in asymptomatic, HIV-1-infected individuals.     

Human coronary transplantation-associated arteriosclerosis. Evidence for a chronic immune reaction to activated graft endothelial cells

Occlusive disease of coronary arteries of engrafted hearts is the major obstacle to long-term survival of human cardiac allografts. The pathogenesis of this process remains uncertain. The identity and localization of cells found in transplantation-associated arteriosclerosis lesions from human cardiac allografts were evaluated, and their expression of class II major histocompatibility complex (human leukocyte antigen-DR [HLA-DR]), surface molecules required for recognition of foreign cells by CD4+ T lymphocytes, was noted. Expanded intimas of transplanted coronary arteries contain T lymphocytes (both CD4+ and CD8+ in approximately equal number) and HLA-DR+ macrophages, both localized primarily in a ring immediately below the luminal endothelium, a distribution strikingly different from that in typical atherosclerosis. Coronary arterial endothelium from six of six transplanted hearts studied bore high levels of HLA-DR. Normal human arteries or usual atherosclerotic lesions have few if any HLA-DR+ endothelial cells. The significance of these findings was tested by evaluating the ability of HLA-DR+ arterial cells to interact with allogeneic T cells in vitro. Endothelial cells (but not smooth muscle cells) cultured from human arteries stimulated foreign CD4+ T cells to proliferate and augmented their secretion of interleukin-2. These findings suggest that ongoing stimulation of recipient T lymphocytes by HLA-DR+ endothelium of donor coronary arteries contributes to a sustained regional immune response. Consequent local release of cytokines may regulate smooth muscle cell proliferation and matrix accumulation within the coronary arteries of allografted hearts.     

Random entry of circulating lymphocyte subsets into peripheral lymph nodes and Peyer's patches: no evidence in vivo of a tissue-specific migration of B and T lymphocytes at the level of high endothelial venules.

Lymphocytes continuously migrate through the body and thus immune competent cells are constantly delivered to most tissues. They interact with high endothelial venules (HEV) via specific homing receptors and vascular addressins, and these molecules seem to be the reason for a preferential homing of B lymphocytes into Peyer's patches and of T lymphocytes into peripheral lymph nodes. When lymphocytes derived from lymph node cell suspensions were applied in the in vitro lymphocyte/endothelium binding assay, the well-known preference of mouse lymph node B lymphocytes for Peyer's patch HEV compared to peripheral lymph node HEV was confirmed in the rat (2.8 times). When in the same in vitro assay thoracic duct lymphocytes (TDL) were used this preference was far less obvious (1.4 times). However, by injecting rat TDL intravenously and by tracing them directly in HEV, B, T, CD4+ and CD8+ lymphocytes are seen to enter Peyer's patches and peripheral lymph nodes in vivo without preference. Thus, in contrast to lymphocytes from lymph node cell suspensions, no evidence was found of a tissue-specific migration of thoracic duct B, T, CD4+ and CD8+ lymphocytes at the HEV level. This finding demonstrates the importance of considering both experimental conditions and the cell source used when investigating lymphocyte traffic.     

Adhesion of peripheral blood lymphocytes of children with arthritis to human umbilical vein endothelial cells.

To determine whether adhesion of peripheral blood lymphocytes (PBL) of patients with juvenile rheumatoid arthritis (JRA) may be enhanced, adhesion of PBL of children with JRA, children with seronegative spondyloarthropathies (SSA), age-appropriate and adult controls, to human umbilical vein endothelial cells (HUVEC) was assessed in vitro. B and CD4 T lymphocytes in initial, adherent, and non-adherent cell fraction were identified by flow cytometry. B lymphocytes of all the younger subjects combined had a higher adherence to activated HUVEC compared with B lymphocytes of the adult donors. Except for greater adherence of HLA-DR+ CD4 T cells, lymphocytes of children with JRA showed no enhanced adhesion to either unactivated or activated HUVEC. The percentage of B cells adherent to activated HUVEC in each of the subject groups was 1.5-3.6-fold higher than adherent CD4 T lymphocytes. Surface analyses indicated higher percentages of CD49d (alpha 4)+ and CD29 (beta 1)+ CD4 T lymphocytes in adherent cells, but less of a differential in CD49 (alpha 4)+ and no difference in CD29 (beta 1)+ B lymphocytes. There were fewer Leu-8 (L-selectin)+ B and Leu-8+ CD4 T cells among adherent cells. The data suggest a greater adhesive capacity of B lymphocytes compared with CD4 T lymphocytes which is unrelated to disease, and the possibility that B lymphocytes may utilize adhesion molecules distinct from those of CD4 T lymphocytes. Only a small subset of T cells of patients with JRA may have an enhanced capacity for adhesion to endothelium.     

Differential expression of beta1 and beta2 integrins and L-selectin on CD4+ and CD8+ T lymphocytes in human blood: comparative analysis between isolated cells, whole blood samples and cryopreserved preparations

Flow cytometric analysis was used to compare the expression of adhesion molecules on human CD4+ and CD8+ T lymphocytes in isolated blood mononuclear cells (MNCs) in whole blood samples and in cryopreserved MNC preparations. Examination of MNCs revealed that the CD11b and CD11c components of the beta2 integrins were preferentially expressed on CD8+ T cells, whereas CD62L was present on more CD4+ T cells. All CD4+ and CD8+ T lymphocytes were positive for CD11a but the CD8+ population had a higher intensity of expression of CD11a and also CD11b. Virtually identical results were obtained with T cells in whole blood samples. In relation to the beta1 integrins, the only difference between isolated CD4+ and CD8+ T cells was that the latter subset had a greater proportion of cells bearing CD49d. The naive cell marker CD45RA was present on the majority of CD8+ T cells whereas CD45RA and the memory marker CD45RO were evenly distributed within the CD4+ T cell subset. Although cryopreservation of lymphocytes did not modify the expression of beta1 and beta2 integrins it produced a marked reduction in the percentage of CD4+ and CD8+ T cells bearing CD62L. With regard to endothelial interactions, it appears that cryopreserved lymphocytes are suitable for inclusion in studies of integrin-mediated adhesion but not for those relating to tethering or recognition of addressins on high endothelial venules. Differences in adhesion molecule expression between CD4+ and CD8+ T lymphocytes could underlie the selective extravasation of these subsets into sites of infection and inflammation.     

The pyrimidin analogue cyclopentenyl cytosine induces alloantigen-specific non-responsiveness of human T lymphocytes

Cyclopentenyl cytosine (CPEC) has been shown to induce apoptosis in human T lymphoblastic cell lines and T cells from leukaemia patients. In this study we have addressed the question of whether CPEC is able to decrease proliferation and effector functions of human alloresponsive T lymphocytes and induce T cell anergy. The proliferative capacity of human peripheral blood mononuclear cells in response to allogeneic stimulation was measured by 5,6-carboxy-succinimidyl-diacetate-fluorescein-ester staining. Flow cytometric analysis was performed using surface CD4, CD8, CD25, CD103 and intracellular perforin, granzyme A, granzyme B, caspase-3 and forkhead box P3 (FoxP3) markers. The in vivo immunosuppressive capacity was tested in a murine skin graft model. Addition of CPEC at a concentration of 20 nM strongly decreased the expansion and cytotoxicity of alloreactive T cells. Specific restimulation in the absence of CPEC showed that the cells became anergic. The drug induced caspase-dependent apoptosis of alloreactive T lymphocytes. Finally, CPEC increased the percentage of CD25(high) FoxP3+ CD4+ and CD103+ CD8+ T cells, and potentiated the effect of rapamycin in increasing the numbers of alloreactive regulatory T cells. Treatment with CPEC of CBA/CA mice transplanted with B10/Br skin grafts significantly prolonged graft survival. We conclude that CPEC inhibits proliferation and cytotoxicity of human alloreactive T cells and induces alloantigen non-responsiveness in vitro.     

Development of lymphocyte subsets in pigtailed macaques

The early development of eight lymphocyte subsets was determined for pigtailed macaque infants from 0 to 800 days of age using two-color flow cytometry and fluorescein- and R-phycoerythrin-conjugated monoclonal antibodies specific for human leukocyte antigens. Four major lymphocyte subsets in monkeys (B, CD4+ T, CD8+ T, and NK cells) could be further divided using two-color analysis. In neonates, the frequency of lymphocyte subpopulations having surface phenotypes found principally on dense, resting cells (IgD+ B cells, Lp220+ CD4+ T cells, and CD18dull CD8+ T cells) was much higher than subpopulations having phenotypes present principally on buoyant, activated cells (IgD- B cells, Lp220- CD4+ T cells, CD18bri CD8+ T cells). There was a complete absence of two CD18bri CD8+ subsets (CD8dull and CD8bri) during the first 300 days of life. The relative proportion of lymphocyte subsets with resting phenotype decreased with increasing age, while the subpopulations associated with activation gradually increased with age. These findings suggest that during early development immunocompetent cells gradually differentiate into activated lymphocytes.     

Reduced lysis by CD8+ cytotoxic T cells in mixed lymphocyte reactions induced via CD4+ T cells exposed to chemically modified antigen presenting cells.

The resistance by T lymphocytes to activation by antigen (anergy) is well documented for CD4+ T-helper (Th) cells, although less is known about CD8+ cytotoxic T lymphocytes (CTL). One widely used method of inducing anergy of CD4+Th is presentation of antigen by ECDI (1-ethyl-3-(3-dimethylamino-propyl)carbodiimide)-fixed antigen-presenting cells (APCs). We report here that in murine mixed lymphocyte reactions (MLRs), a marked reduction in detected cytotoxicity (which is mediated predominantly by CD8+ CTL) occurs on day 7 if the bulk cultures are restimulated 2 days previously with ECDI-fixed allogeneic splenocytes. No differences were seen between untreated cultures on days 5 and 7, or on day 7 of cultures to which were added unfixed allogeneic splenocytes, fixed or unfixed syngeneic splenocytes, or 'third-party' allogeneic splenocytes, 2 days previously. The effect is not mediated directly on CD8+ cells, since MLRs depleted of CD4+ cells immediately prior to exposure to fixed allogeneic splenocytes fail to show reduced lysis. On the other hand, reduced lysis did occur if CD4+ cells, purified from the MLRs on day 4, were exposed to ECDI-fixed allogeneic splenocytes and then returned to MLRs previously depleted of CD4+ cells. Moreover the effect is overcome using exogenous interleukin-2 (IL-2). We propose that CD4+ cells, restimulated by a regimen shown previously to induce their anergy, can cause a reduction in CD(8+)-mediated cytotoxicity in MLRs.     

Both LFA-1-positive and -deficient T cell clones require the CD2/LFA-3 interaction for specific cytolytic activation.

We investigated the capacity of T lymphocytes from a leukocyte adhesion-deficient (LAD) patient to respond to alloantigen. Leukocytes of this patient completely lacked LFA-1 surface expression due to the absence of mRNA coding for the LFA-1 beta chain. Despite the absence of LFA-1, T lymphocytes obtained from this patient, cultured with allogeneic stimulator cells (lymphoblastoid B cells JY), were capable of lysing JY cells. Furthermore, two T cell clones (one CD4+ and one CD8+), generated from this lymphocyte culture, specifically lysed the allogeneic lymphoblastoid JY cells. The cytolytic capacity of LFA-1-negative T lymphocytes and T cell clones was comparable to that of control LFA-1-positive T cells with allospecificity against JY. Detailed analysis of the CD4 positive and LFA-1-negative T cell clone demonstrated that it specifically recognized HLA-DQ. Antibody inhibition studies showed that the CTL/target cell interaction was mediated through the CD2/LFA-3 adhesion pathway. LFA-1 expressed by the target cells did not participate in the CTL/target cell conjugate formation and contributed only minimally to the cytotoxic activity. Moreover, when allogeneic LFA-1-deficient B cells, bearing the appropriate HLA-DQ alloantigen, were used as target cells, significant levels of specific cytotoxicity were measured, further excluding a role for LFA-1 in this interaction. The adhesion molecules, VLA-4, CD44 and L-selectin (LECAM1) were not involved. These results demonstrate that LFA-1-negative T lymphocytes can exert allospecific cytotoxicity and that CTL/target cell contact is mediated through the CD2/LFA-3 route. This observation may explain in part why in LAD patients viral infections, cleared largely by T cells, are less frequently observed than bacterial infections, in which phagocytic cells play a major role.     

Effects of mibefradil, a selective T-type Ca2+ channel antagonist, on sino-atrial node and ventricular myocardia

The effects of mibefradil, a non-dihydropyridine Ca2+ channel antagonist, on the action potential configuration of isolated rabbit sino-atrial node preparations, membrane currents of guinea-pig ventricular myocytes and the contractile force of isolated ventricular papillary muscles were examined. In sino-atrial node preparations, 10 microM mibefradil decreased the slope of the pacemaker depolarization (phase 4 depolarization) and maximum rate of rise, and shifted the threshold potential to the positive direction with no effect on action potential duration. In ventricular myocytes, 1 microM mibefradil inhibited the T-type Ca2+ current by about 40% while it had no effect on the L-type Ca2+ current. At 10 microM, mibefradil inhibited the L-type and T-type Ca2+ currents by about 40% and 90%, respectively. Mibefradil had no effect on contractile force at concentrations up to 1 microM. Thus, mibefradil was shown to produce potent prolongation of the pacemaker depolarization, mainly through inhibition of the T-type Ca2+ current. It is suggested that the T-type Ca2+ current may not be involved in ventricular contraction.     

Functional analysis of mononuclear cells infiltrating into tumors. VI. The effect of lymphocyte chemotactic factors on lymphocyte adhesion to endothelial cells.

We have analyzed mechanisms controlling infiltration of T lymphocytes into tumor tissues. A lymphocyte chemotactic factor-b (LMF-b) produced by tumor infiltrating CD4+ T lymphocytes was purified. LMB-b was specifically chemotactic for CD8+ T lymphocyte. Furthermore, LMF-b augmented lymphocyte adhesion to high endothelial venule (HEV) cells. The binding of CD8+ T cells to HEV cells was specifically augmented by LMF-b. The LMF-b primarily acted on T lymphocytes, whereas tumor necrosis factor as well as IFN-gamma acted on HEV cells or fibroblast cells. The binding of lymphocytes to fibroblast cell line was not augmented by LMF-b. The augmentation of lymphocyte adhesion to endothelial cells by LMF-b was mediated by the lymphocyte function associated antigen-1/intercellular adhesion molecule (LFA-1/ICAM) pathway, the CD2/LFA-3 pathway, and the very late antigen-4/culture supernatant-1 (VLA-4/CS-1) pathway.     

Interleukin-7 is a potent co-stimulus of the adhesion pathway involving CD2 and CD28 molecules.

Co-stimulation of highly purified peripheral T lymphocytes from healthy blood donors with the adhesion molecules CD2 and CD28 in association with recombinant interleukin-7 (rIL-7) induced T-cell proliferation, multiple cytokine secretion and IL-2 receptivity. We demonstrated that rIL-7 is as potent as rIL-2 in inducing the proliferation of unseparated, CD4+ and CD8+ T cells. In contrast to low or undetectable levels of IL-1 alpha, IL-6 and IL-2, high levels of tumour necrosis factor-alpha (TNF-alpha), IL-4 and granulocyte-macrophage colony-stimulating factor (GM-CSF) were secreted. Experiments using blocking antibodies suggested a direct mechanism for rIL-7 co-stimulatory effect, although induction of the CD25/IL-2 receptor alpha-chain (CD25/IL-2R alpha) was observed. Monoclonal antibodies (mAb) against the adhesion molecules CD2 and CD28 are likely to mimic the interaction with their respective physiological ligands [lymphocyte function-associated antigen-3 (LFA-3)/CD58, CD59 and CD48 for CD2, B7/BB1 for CD28]. Taken together, these in vitro data suggest that IL-7 could participate in paracrine interactions between T lymphocytes and thymic stromal cells or dendritic cells, via its potent co-stimulatory activity with CD2 and CD28 adhesion molecules.     

Unusual expression of CD2 in sheep: implications for T cell interactions

The CD2 adhesion/activation molecule on the surface of mammalian T lymphocytes binds to a ubiquitous receptor, LFA-3. We show that CD2 in sheep differs significantly in its expression from CD2 in humans, and this most likely relates to the high level of expression of the sheep LFA-3 molecule. In sheep, in contrast to man, CD2 was weakly expressed on peripheral T cells and thymocytes. Moreover, a large subset of T cells identified by the monoclonal antibody T19 and considered to be gamma/delta receptor-bearing T cells completely lacked the CD2 molecule. T19+ cells constituted up to 50% of peripheral blood T cells in lambs, and 20-30% of T cells in older sheep, whereas the CD4+ and CD8+ subsets, which are both CD2+, constituted relatively small subsets in peripheral blood. Only those T cells which did express CD2 adhered as "rosettes" to dendritic cells, and the localization of CD2 to the membrane junction indicated that CD2 was critical for this adhesion. However, CD2 adhesion was not necessary for CTL-mediated killing of allogeneic target cells, since T19+ cells generated in bulk mixed lymphocyte culture were extremely efficient at killing appropriate target cells. Some of the behavioral differences between T19+ and CD4+/CD8+ subsets might be explained by the presence or absence of CD2. The results also indicate that the expression of CD2 (and LFA-3) may differ markedly between species.     

Enhanced adhesion of autologous lymphocytes to gamma-interferon-treated human endothelial cells in vitro.

An adhesion assay was developed using human umbilical vein endothelial cell cultures and autologous and allogeneic lymphocytes separated from cord blood. Endothelial monolayers cultured in plain or gamma-interferon (IFN gamma)-supplemented medium were cocultured with lymphocytes for 1 h and non-adherent lymphocytes removed by washing. Autologous and allogeneic lymphocytes exhibited significantly increased adhesion with IFN gamma-treated cells compared with untreated controls. The increased adhesion to IFN gamma-treated endothelial cells was significantly inhibited when autologous or allogeneic lymphocytes were pre-treated with saturating amounts of an anti-CD4 monoclonal antibody. The results indicate that IFN gamma enhances lymphocyte binding to endothelium and that the CD4 molecule may be involved in this process. This could be an important mechanism in targetting the migrating of T-helper (Th) cells to areas of chronic inflammation and in antigen presentation by endothelial cells.     

The effect of cytokines and chemotactic agonists on the migration of T lymphocytes into skin.

The migration of lymphocytes and neutrophils into skin sites stimulated with chemotactic agonists or with cytokines known to induce leucocyte-endothelial adhesion molecules was examined in sheep. Lymphocytes, collected from efferent prefemoral lymph, labelled in vitro with [111In]oxine and reinjected intravenously, migrated in large numbers into delayed-type hypersensitivity (DTH) reactions elicited by purified protein derivative (PPD) and into sites stimulated with tumour necrosis factor-alpha (TNF-alpha) or interferon-gamma (IFN-gamma). In contrast, interleukin (IL)-1 alpha, a potent inducer of endothelial leucocyte adhesion molecule-1 (ELAM-1), caused moderate accumulation of [111In]lymphocytes at concentrations that induced intense accumulation of 111In-labelled neutrophils. The chemotactic agonists IL-8 and zymosan-activated plasma (ZAP) caused accumulation of very large numbers of neutrophils but only small numbers of lymphocytes, whereas platelet-activating factor (PAF) and leukotriene B4 (LTB4) failed to recruit lymphocytes into skin. IFN-gamma was the only mediator to recruit lymphocytes in preference to neutrophils into skin. The results suggest that those lymphocyte chemotactic agonists which lack the ability to induce adhesion molecules on endothelium play only a minor role in directing migration of lymphocytes into skin. Immunohistological examination of skin lesions confirmed the findings of studies with 111In-labelled lymphocytes and indicated that there was a tendency for CD4+ cells to outnumber CD8+ cells in infiltrates induced by all mediators. In contrast to the elevated numbers of T19+ subset of T-cell receptor (TcR+) gamma delta cells present in DTH reactions, none of the mediators induced migration of T19+ cells into skin.     

Vulnerable caregivers of patients with Alzheimer's disease have a deficit in circulating CD62L- T lymphocytes

OBJECTIVE: The cell adhesion molecule, L-selectin (CD62L), serves a crucial role in the migration of naive T lymphocytes and is typically shed on cell activation. The objective of this study was to determine the effects of chronic stress on L-selectin expression on peripheral lymphocytes in elderly spousal caregivers of patients with Alzheimer's disease. METHODS: Twenty caregivers (mean age, 73.5 years) had their lymphocytes and catecholamine levels sampled at rest and in response to an acute psychological stressor. Ten of the caregivers were classified as susceptible or "vulnerable" based on the large amount of care required by the patient relative to the amount of respite the caregiver received during the previous 6 months. RESULTS: At rest, vulnerable caregivers had 60% fewer L-selectin negative CD8+ T cells (CD8+CD62L-) (p=.01) but no difference in CD8+CD62L+ cells. Vulnerable caregivers also showed significantly fewer CD4+CD62L- T lymphocytes (p=.04) but no difference in CD4+CD62L+ lymphocytes. Resting plasma epinephrine levels were 44% higher in vulnerable caregivers as compared with nonvulnerable caregivers (p=.01). The acute stressor increased circulating levels of CD8+CD62L- and CD8+CD62L+ lymphocytes and catecholamines similarly in both groups. CONCLUSIONS: The findings suggest that caregivers who are more vulnerable to the chronic stress of caregiving show a decrement in circulating CD62L- T lymphocytes, possibly by adrenomedullary activation. The data also suggest the identity of lymphocyte subsets that may underlie prior observations of immunologic decrements associated with the chronic stress of caregiving.     

Application of anti-CD3-treated lymphocytes as stimulator cells in human autologous mixed lymphocyte cultures for generation of autorestricted CD8+ suppressor T cells.

The purpose of the present study was to investigate the application of anti-CD3-treated lymphocytes as stimulator cells in human one-way autologous mixed lymphocyte cultures (MLC) for the generation of suppressor cells. MLC-activated CD4-CD8+ CD16- T cells were non-cytotoxic, while they down-regulated the proliferation of autologous (but not allogeneic) responder lymphocytes in allogeneic test MLC.