Prevention of cyclophosphamide-induced and spontaneous diabetes in NOD/Shi/Kbe mice by anti-MHC class I Kd monoclonal antibody

The immune mechanisms directly responsible for beta-cell destruction in insulin-dependent diabetes are undefined. We studied the role of MHC class I-restricted T lymphocytes in the development of diabetes in cyclophosphamide (CY)-treated male and untreated female NOD mice (H-2Kd,Db). After administration of CY to 10-wk-old male NOD/Shi/Kbe mice, 37 of 64 (58%) phosphate-buffered saline-injected control mice and 13 of 22 (59%) anti-Kb and 12 of 27 (44%) anti-Db monoclonal antibody (MoAb)-injected mice became diabetic by 14 wk of age, whereas only 3 of 38 (8%) anti-Kd and 2 of 13 (15%) anti-Lyt-2 MoAb-injected mice did. In untreated female NOD/Shi/Kbe mice, 30 of 46 (65%) mice developed spontaneous diabetes by 30 wk of age, whereas none of 9 anti-Kd MoAb-injected mice became diabetic. Immunohistochemical studies showed that islet-infiltrating cells in CY-treated control mice were composed mainly of both L3T4+ and Lyt-2+ T lymphocytes, whereas many L3T4+ and very few Lyt-2+ lymphocytes infiltrated within the islets in anti-Kd MoAb-injected mice. Administration of anti-Lyt-2 MoAb induced the absence of Lyt-2+ T lymphocytes in the islet and spleen. However, anti-Kd MoAb did not change the number of spleen cells or the T-lymphocyte subset and response to concanavalin A. These results suggest that MHC class I Kd-restricted Lyt-2+ T lymphocytes play an important role as direct effector cells in destruction of beta-cells in NOD/Shi/Kbe mice.     

The effects of immunosuppressants on FAS-mediated activation-induced cell death in human T lymphocytes

The effects of cyclosporin A (CsA) and methylprednisolone (MP) on Fas-mediated activation-induced cell death (FMAICD) of T lymphocytes were examined. T lymphocytes were activated with the immobilized anti-CD 3 and CD 28 monoclonal antibodies (MoAbs) (activation phase) and incubated further with the agonistic MoAb against Fas (death phase). Cell proliferation and DNA fragmentation were measured by XTT and diphenylamine assay. CsA in the activation phase inhibited DNA fragmentation mediated by anti-Fas MoAb but not MP. The combination of CsA and MP at the lower concentrations had little effect on FMAICD, although they had similar degrees of suppression on T lymphocyte proliferation as the maximum obtained by CsA or MP alone. In the death phase, MP induced apoptosis without 7C11 and CsA had no effects. These results indicate that the combination of CsA and MP at low concentrations could maintain FMAICD with the suppression on T lymphocyte proliferation.     

Specific binding and lysis of human melanoma by IL-2-activated cells coated with anti-T3 or anti-Fc receptor cross-linked to antimelanoma antibody: a possible approach to the immunotherapy of human tumors

Monoclonal antibodies (MoAb) to human melanoma have demonstrated a limited ability to cause tumor regression in humans when used alone or when coupled to gamma-emitting radioisotopes. We have evaluated heteroaggregates containing antilymphocyte antibodies crosslinked to antimelanoma monoclonal antibodies recognizing p97, a transferrin-like molecule (MoAb 96.5). When coupled to antibodies recognizing T3 (CD3, part of the T-cell receptor complex for antigen) or to 3G8, an antibody recognizing the Fc receptor present on large granular lymphocytes and granulocytes (CD16), significant induction of effector target crosslinks and target cell lysis could be obtained. Effector cells incubated for 24 hr with recombinant IL-2 were coated with the crosslinked reagents and tested for conjugate formation and for cytotoxicity in a 4-hr assay with chromium-labeled targets. Marked increases in conjugation to autologous tumor (47.0% compared to 11.8%) was demonstrated with E+ cells using the T3-coupled MoAb and with E- cells using the Fc receptor-coupled MoAb (22.6% compared to 11.2%). When tested in sequential cytotoxicity assays using unseparated effector cells incubated for 1, 2, and 3 days in IL-2, lytic activity was less than 2, less than 2, and 3.3 LU/10(6) cells for cells incubated in monomeric 96.5; 2.6, 5.3, and 50 LU/10(6) cells incubated in 96.5 crosslinked T3; and less than 2, 3.6, and 8.0 LU/10(6) cells for cells incubated with 96.5 crosslinked to 3G8. Similar findings were noted in two other experiments. Heteroaggregates such as these may be useful in conjunction with the transfer of IL-2-activated cells or with IL-2 alone in immunotherapy trials.     

The response of human T lymphocytes against alloantigens following Fas-mediated activation-induced cell death

We examined the response of T lymphocytes activated with specific alloantigens following Fas-mediated apoptosis; using a mixed lymphocyte culture (MLC) system. Cells obtained from an MLC after 6 or 7 days of culture were incubated for are additional 24 hours in the presence or absence of the agonistic monoclonal antibody (MoAb), 7C11, or the antagonistic MoAb, ZB4. We assessed DNA fragmentation/specific cytotoxiy of the MoAb-treated cells. Cells harvested after 4 days of culture were sensitive to apoptosis induced by 7C11 with maximum DNA fragmentation observed on day 6. ZB4 slightly inhibited apoptosis of the cells compared with controls. The simultaneous addition of recombinant interleukin-2 (rIL-2) with the MoAbs significantly inhibited DNA fragmentation in control and ZB4-treated cells, but had little effect on the 7C11-treated cells. Control and ZB4-treated MLC cells showed cytotoxic activities against specific target cells, namely >10%. In contrast, the 7C11-treated cells showed <5% cytotoxicity. Although the addition of rIL-2 increased specific percentage cytotoxicity of control and ZB4-treated cells, it had little effect on the specific cytotoxic activity of the 7C11-treated MLC cells. These results suggest that specific cytotoxic T lymphocytes may be eliminated via apoptosis mediated by the Fas/Fas ligand system.     

The effect of anti-interleukin 2 monoclonal antibody treatment on the survival of rat cardiac allograft

The effect of anti-interleukin 2 monoclonal antibody (anti-IL2 MoAb) and the accumulation of intravenously administered 125I-labeled anti-IL2 MoAb were examined in heterotopic rat cardiac allografts. Mouse anti-human recombinant IL2 MoAb was obtained by the hybridoma technique. The anti-IL2 MoAb, termed 8H-10, was an IgG2a which inhibited IL2-driven [3H]TdR incorporation in cytolytic T lymphocyte line cells at a dilution of 2(6). 8H-10 was injected iv at a dose of 200 micrograms/day for 8 consecutive days, beginning on the day of transplantation. Hearts from F344 rats (RT11v1) were transplanted into ACI recipient rats (RT1av1). The mean survival time was 7.6 +/- 0.8 days in untreated controls, 9.0 +/- 1.2 days in additional controls treated with mouse anti-sheep red blood cell monoclonal antibody, and 25.3 +/- 18.4 days in the anti-IL2 MoAb (8H-10)-treated group (P less than 0.05). Furthermore, the accumulation of intravenously administered 125I-labeled anti-IL2 MoAb (8H-10) was specifically seen in the grafted heart. In conclusion, these results suggest that IL2 may play an important role in allograft rejection and that anti-IL2 MoAb may serve as a useful immunosuppressive agent in clinical transplantation.     

Immunomagnetic T-Lymphocyte Depletion (ITLD) of Rat Bone Marrow Using OX-19 Monoclonal Antibody

Graft versus host disease (GVHD) may be abrogated and host survival prolonged by in vitro depletion of T lymphocytes from bone marrow (BM) prior to allotransplantation. Using a mouse anti-rat pan T-lymphocyte monoclonal antibody (0×19) bound to monosized, magnetic, polymer beads, T lymphocytes were removed in vitro from normal bone marrow. The removal of the T lymphocytes was confirmed by flow cytometry. Injection of the T-lymphocyte-depleted bone marrow into fully allogeneic rats prevents the induction of GVHD and prolongs host survival.

A highly efficient technique of T-lymphocyte depletion using rat bone marrow is described. It involves the binding of OX-19, a MoAb directed against all rat thy-mocytes and mature peripheral T lymphocytes, to monosized, magnetic polymer spheres. Magnetic separation of T lymphocytes after mixing the allogeneic bone marrow with the bead/OX-19 complex provides for a simple, rapid depletion of T lymphocytes from the bone marrow. In vitro studies using flow cytometry and the prevention of GVHD in a fully allogeneic rat bone marrow model have been used to demonstrate the effectiveness of the depletion procedure.     

Soluble CD8, IL-2 receptor, and tumor necrosis factor-alpha levels in steroid-resistant acute graft-versus-host disease. Relation with subsequent response to anti-IL-2 receptor monoclonal antibody treatment

Serial determination of soluble CD8 (sCD8), soluble IL-2 receptors (sIL-2R), and tumor necrosis factor-alpha serum levels were performed in bone marrow transplant patients upon initiation, day 0 (D0) and at D10 of an anti-IL-2 receptor (alpha chain) monoclonal antibody (B-B10) in vivo treatment for steroid-resistant grade greater than or equal to 2 acute graft-versus-host disease (aGVHD). D0 and D10 sCD8 serum levels correlated strongly with response to B-B10 treatment (p = .003 and .001, respectively); 76% of the patients with D0 sCD8 levels less than 500 U/ml responded favorably to B-B10 treatment, versus only a 30% response if the sCD8 levels were greater than 500 U/ml (p = .02). Likewise, D0 tumor necrosis factor-alpha levels significantly correlated with subsequent response to B-B10 treatment (p = .03). D0 sIL-2R levels were not significantly different in B-B10-responsive and nonresponsive aGVHD patients. These results suggest that the serial determination of sCD8 and TNF serum levels could provide valuable predictive information as to steroid-resistant aGVHD responsiveness to anti-IL-2R treatment.     

Expression of Ly-6C by T lymphocytes of NOD mice after CD3-complex stimulation. Identification of activated cells during insulitis of prediabetic mice

Ly-6C is a differentiation antigen that distinguishes T-lymphocyte subsets. In concordance with previous results, splenocytes from NOD mice do not express the epitope recognized by anti-Ly-6C monoclonal antibodies (MoAbs), including MoAb HK1.4 in this study, and cannot be stimulated to proliferate in response to HK1.4. However, when splenocytes from NOD mice were stimulated in vitro with the anti-CD3 MoAb 145-2C11, T lymphocytes expressing Ly-6C were detected after 48 h of stimulation, with as many as 25% of lymphocytes expressing this antigen with prolonged passage in culture. Most of the cells expressing Ly-6C were Thy-1.2+, CD4+, and CD8- and proliferated after stimulation with HK1.4. To further understand the failure of NOD splenocytes to express Ly-6C, freshly isolated cells were stimulated with alpha/beta-interferon (IFN-alpha/beta) and IFN-gamma. Although these lymphokines induced expression of Ly-6A and Ly-6C in splenocytes from C57BL/6J mice and Ly-6A in NOD cells, Ly-6C was not induced on NOD cells. Because Ly-6C expression on splenocytes was a marker of activation via the CD3 T-lymphocyte receptor complex, we also examined expression of Ly-6C on T lymphocytes within islets showing insulitis in vivo. Lymphocytes that were Ly-6C+ were identified within islets on histological sections of pancreas, whereas Ly-6C+ cells in the spleen from the same mouse could not be detected. Our findings imply functional abnormality in expression of Ly-6C in NOD mice.(ABSTRACT TRUNCATED AT 250 WORDS)     

Anti-interleukin 2 receptor monoclonal antibody in the treatment of ongoing acute rejection episodes of human kidney graft--a pilot study

Monoclonal antibodies (MoAbs) against human interleukin 2 receptor (IL-2-R) have been shown to prevent early kidney rejection in animals and humans. We report here the effect of an anti-IL-2-R MoAb (33B3.1) inhibiting IL-2 binding high-affinity sites on activated lymphocytes in 10 declared acute rejection episodes of first cadaveric kidney grafts. Six patients were under cyclosporine treatment only at the time of diagnosis of the rejection. All rejection episodes but one were biopsy-proved cellular rejections. Treatment consisted of intravenous infusions of 33B3.1 at 20 mg/day x 2 days, followed by 10 mg/day for 8 additional days. In case of MoAb ineffectiveness at day 5, anti-IL-2-R MoAb was discontinued and a rescue treatment of corticosteroid boluses (CSb) was given. If not, in all cases corticosteroids (CS) were given (1 mg/kg) at the end of MoAb treatment (day 10) and tapered off thereafter. Two rejection episodes immediately responded to 33B3.1 treatment. During 33B3.1 treatment four other patients had only a stabilization of their blood creatinine concentration, which nevertheless returned to prerejection levels after day 10 when anti-IL-2-R was discontinued and CS administered at 1 mg/kg (no rescue treatment). The four remaining patients had an increase of their blood creatinin levels at day 5 despite 33B3.1 treatment, and their renal function only improved with CSb rescue treatment. One of these patients lost the graft despite rescue treatment, as well as a 9-day course of antithymocyte globulin. Trough levels of MoAb reached a plateau as early as day 2 (approximately 6 micrograms/ml). All patients developed antibodies (IgM and IgG) after day 14. In no instance could unresponsiveness be related to low circulating 33B3.1 trough levels or to early host anti-MoAb immune response (IgM or IgG). We conclude that 33B3.1, known to be effective in preventing early rejection, has only inconsistent and/or incomplete effects on the ongoing rejection process. Our data suggest that once IL-2-dependent clones are expanded in the rejected graft, interference with IL-2/IL-2-R signals does not block the effector mechanisms sustaining acute rejection.     

Expression of the interleukin 2 receptor beta chain (p75) in renal transplantation--applicability of anti-interleukin-2 receptor beta chain monoclonal antibody.

The interaction of interleukin 2 with specific cellular receptors plays an essential role in the allostimulated proliferation and differentiation of T cells. Recent chemical linking studies have demonstrated that the human high-affinity IL-2 receptor is a membrane complex composed of at least two distinct subunits, which are the p55 (alpha-chain) and p75 (beta-chain) subunits. The IL-2R beta chain is supposed to play a role in the signal transduction of IL-2, but the exact mechanism is still unknown. In this study, we investigated the effects of a newly established anti-IL-2R beta chain monoclonal antibody (MoAb, TU-27) on the induction of cytotoxic T lymphocytes (CTLs) using the cell-mediated lympholysis (CML) assay. TU-27 in combination with H-31, a MoAb directed against the IL-2R alpha chain, produced inhibition of cytotoxicity, while TU-27 alone could not inhibit cytotoxicity, while TU-27 alone could not inhibit cytotoxicity at any concentration. TU-27 plus H-31 prevented the expansion of CD4+ cells and CD8++ cells in mixed lymphocyte culture (MLC). Furthermore, we examined the serial changes in the expression of the IL-2R beta chain on peripheral blood lymphocytes from renal transplant recipients using two-color immunofluorescence flow cytometry, so as to investigate correlations between IL-2R beta chain expression and the occurrence of allograft rejection. Here, we report that the IL-2R beta chain is expressed on CD4-positive (CD4+) cells and strongly CD8-positive (CD8(+)+) cells in association with acute rejection, indicating that IL-2R beta chain expression appears to increase on alloreactive T cells.     

Prevention of autoimmune diabetes with nonactivating anti-CD3 monoclonal antibody.

Autoreactive T cells mediate diabetes in animal models of insulin-dependent diabetes mellitus (IDDM) and are believed to cause the disease in humans. Therefore, immunotherapies directed against T cells are of particular interest for the treatment of IDDM. One candidate for such immunotherapy is anti-CD3 monoclonal antibodies (MoAbs), but clinical side effects are common with anti-CD3 treatment due to the ability of these MoAbs to activate T cells in vivo. However, F(ab')2 fragments of anti-CD3 are nonactivating and immunosuppressive. We evaluated the effects of whole anti-CD3 MoAb and F(ab')2 fragments in the setting of experimental autoimmune diabetes. Treatment with whole MoAb or F(ab')2 fragments significantly reduced the hyperglycemia induced with multiple low dosages of streptozocin (MDSDM; 232 +/- 23 mg/dl, P less than 0.01 and 235 +/- 16 mg/dl, P less than 0.01 vs. 325 +/- 25 mg/dl, respectively) in male CD1 mice. Both whole MoAb and F(ab')2 fragments suppressed the development of insulitis (P less than 0.001). Treatment with whole MoAb resulted in marked weight loss (10.4 +/- 1.5% of total body wt), and the mice appeared ill and listless, whereas, mice treated with F(ab')2 fragments gained weight (4.9 +/- 5.5% of total body wt) and appeared healthy. Treatment with whole MoAb caused activation of T cells in vivo as reflected by proliferation of freshly isolated spleen cells to recombinant interleukin-2. Depletion of T cells with whole MoAb was more pronounced than with F(ab')2 fragments, and T-cell receptor (TCR) reexpression on remaining cells occurred with F(ab')2 fragments within 48 h after F(ab')2 treatment.(ABSTRACT TRUNCATED AT 250 WORDS)     

Combination anti-CD2 and anti-CD3 monoclonal antibodies induce tolerance while altering interleukin-2, interleukin-4, tumor necrosis factor, and transforming growth factor-beta production.

OBJECTIVE: These studies were designed to elucidate the mechanism by which signals delivered by anti-CD2 monoclonal antibody (MoAb) interfere with activational signals delivered by anti-CD3 MoAb and induce long-term graft survival and tolerance. SUMMARY BACKGROUND DATA: Anti-CD2 or anti-CD3 MoAb can prolong allograft survival when administered alone. In combination, they synergistically prolong survival while reducing anti-CD3-associated cytokine toxicity. It was postulated that the mechanism of synergism and reduced cytokine toxicity was related to anti-CD2-induced alterations in anti-CD3-induced T-cell activation. METHODS: C57BL/6 (H-2b) mouse hearts were transplanted to CBA (H-2k) mice. The recipients received anti-CD2 and/or anti-CD3 MoAb intravenously only at the time of initial allografting. Serum from treated animals and culture supernatants from lymphocytes stimulated in vitro with anti-CD3 were examined for interleukin (IL)-2, -4, -6, and -10, tumor necrosis factor (TNF), and transforming growth factor-beta (TGF beta). RNA was isolated from lymphocytes from treated animals and examined for receptor and cytokine gene expression by northern hybridization or reverse transcribed and amplified by the polymerase chain reaction (PCR). RESULTS: Anti-CD2 and anti-CD3 MoAbs alone prolonged graft survival (22.0 +/- 0.5 days and 28.0 +/- 0.5 days, respectively; p < 0.02 and p < 0.01 vs. control, by Wilcoxon signed-rank test). Combined anti-CD2/anti-CD3 MoAbs synergistically prolonged survival indefinitely (> 150 days, p < 0.01) while decreasing cytokine toxicity. Second donor-specific allografts also showed long-term survival. The peak serum TNF concentration (2100 units/mL) was reduced 78% by anti-CD2 treatment (455 units/mL). Anti-CD2 inhibited anti-CD3-stimulated proliferation and in vitro production of IL-2 and IL-4, with no alteration of IL-6, IL-10, or TNF. Conversely, there was an increase in the immunosuppressive cytokine TGF beta. PCR analysis showed that anti-CD2 reduced anti-CD3-stimulated IL-2 messenger RNA expression, and by northern analysis, anti-CD2 inhibited anti-CD3-stimulated increases in messenger RNA for the CD2 and CD3 receptors themselves. CONCLUSIONS: The combination of anti-CD2 and anti-CD3 MoAbs induced a state of tolerance while decreasing anti-CD3-associated cytokine toxicity. The mechanism was related to anti-CD2-generated alterations in T-cell activation and gene expression.     

Characterization of immobilized anti-CD3 antibody-activated T lymphocytes for use in adoptive immunotherapy of patients with brain tumors.

Large numbers of T lymphocytes were successfully cultured for use in adoptive immunotherapy using immobilized anti-CD3 antibody. This method allows more than 1.5 x 10(10) T lymphocytes to be cultured within 2 weeks from only 20 ml of blood. Proliferated T lymphocytes were less cytotoxic than lymphokine-activated killer cells induced with a high dose of interleukin-2, but were more specific for autologous tumor cells as determined by cold target competition. Signal transduction of anti-CD3 antibody induced a population of CD8 positive cells, i.e. cytotoxic T lymphocytes. Adoptive immunotherapy using autologous lymphocytes requires a sufficient number of lymphocytes and high induced cytotoxic activity. This method is promising for clinical adoptive immunotherapy of patients with brain tumors.     

Effect of Surgery on the Quantity of Lymphocyte Subpopulations.

Surgical procedures cause an equal decrease in the number of peripheral blood T and B lymphocytes. This decrease attained its lowest value in 2 to 7 days with recovery or approaching recovery by 10 days. The percentages of T and B lymphocytes obtained postoperatively do not vary from preoperative values; this indicates an equal depression of lymphocytes rather than selective depression of one of the two lymphocyte subpopulations. Neutrophils increased and eosinophils decreased, attaining average maximum and minimum values, respectively, at 2 days postoperatively with subsequent recovery by 10 days.     

微波烘疗对原发性淋巴水肿免疫细胞的影响

目的　探明原发性淋巴水肿患者免疫细胞之变化情况。方法　采用 Ａ Ｂ Ｃ 及 Ａ Ｐ Ａ Ａ Ｐ法免疫组织化学技术,分别对１０ 例原发性下肢淋巴水肿患者患肢皮肤及外周血中淋巴细胞等进行分类分析,并观察微波烘疗的影响。结果　外周血 Ｃ Ｄ８ ＋ Ｔ 淋巴细胞增加、 Ｃ Ｄ４ ＋ Ｔ 淋巴细胞降低, Ｃ Ｄ４／ Ｃ Ｄ８ 降低;局部皮肤真皮层小血管、毛细血管周围可见显著的呈灶性分布的单个核细胞浸润,以 Ｔ 淋巴细胞和单核巨噬细胞为主。结论　微波烘疗后可提高外周血中 Ｃ Ｄ４ ＋ Ｔ 淋巴细胞、降低 Ｃ Ｄ８ ＋ Ｔ 淋巴细胞,恢复 Ｃ Ｄ４／ Ｃ Ｄ８ 比值,因而可调节存在的免疫紊乱状态;对局部组织可显著降低 Ｔ 淋巴细胞浸润,提高巨噬细胞活性,增强其吞噬能力。认为微波烘疗通过其热效应及复杂的生物学效应,纠正原发性淋巴水肿患者全身免疫紊乱状态,促进局部组织炎症反应消褪及增强巨噬细胞吞噬作用,从而达到消除水肿、减轻角质细胞增生及纤维化形成的作用。     

The effect of cyclosporine on ornithine decarboxylase induction with mitogens, antigens, and lymphokines

Ornithine decarboxylase (ODC) is the initial enzyme in polyamine synthesis. An increase in ODC activity is associated with increased RNA, DNA, and protein synthesis. We have used the induction of ODC by mitogens and alloantigens in human peripheral blood lymphocytes as an intracellular marker of protein synthesis and lymphocyte activation. The immunosuppressive agent cyclosporine was found to inhibit both the mitogen and alloantigen stimulated induction of ODC in lymphocytes in a manner that parallels inhibition of subsequent 3H-thymidine incorporation. When purified T lymphocytes were stimulated with mitogen alone, minimal ODC activity was detected. The addition of 5% monocytes, human Interleukin-1 (IL-1), or T cell growth factor (IL-2) enhanced mitogen-induced ODC activity in T lymphocytes 4-10-fold. Cyclosporine inhibited the induction of ODC when T lymphocytes were combined with monocytes or growth factors. We conclude that (1) the induction of ODC in human lymphocytes by mitogen and alloantigen is inhibited in the presence of cyclosporine; (2) the induction of ODC activity in purified T lymphocytes requires the presence of both mitogen and monocytes or their products; (3) IL-1 and IL-2 can supplement for monocytes and augment the phytohemagglutinin induction of ODC in T lymphocytes; and (4) cyclosporine inhibits ODC induction in T lymphocytes stimulated with mitogen in the added presence of monocytes, IL-1, or IL-2. The inhibition of ODC induction and polyamine synthesis by cyclosporine adds insight into its mode of action on the mechanisms involved in early T cell activation.     

微波烘疗对原发性淋巴水肿免疫细胞的影响

目的探明原发性淋巴水肿患者免疫细胞之变化情况。方法采用 ABC 及 APAAP法免疫组织化学技术,分别对10例原发性下肢淋巴水肿患者患肢皮肤及外周血中淋巴细胞等进行分类分析,并观察微波烘疗的影响。结果外周血 CD8 T 淋巴细胞增加、CD4 T 淋巴细胞降低,CD4/CD8降低;局部皮肤真皮层小血管、毛细血管周围可见显著的呈灶性分布的单个核细胞浸润,以 T 淋巴细胞和单核巨噬细胞为主。结论微波烘疗后可提高外周血中 CD4 T 淋巴细胞、降低CD8 T 淋巴细胞,恢复 CD4/CD8比值,因而可调节存在的免疫紊乱状态;对局部组织可显著降低T淋巴细胞浸润,提高巨噬细胞活性,增强其吞噬能力。认为微波烘疗通过其热效应及复杂的生物学效应,纠正原发性淋巴水肿患者全身免疫紊乱状态,促进局部组织炎症反应消褪及增强巨噬细胞吞噬作用,从而达到消除水肿、减轻角质细胞增生及纤维化形成的作用。     

Allo-specific T-Cells Encoding for Viral IL-10 Exert Strong Immunomodulatory Effects in vitro but Fail to Prevent Graft Rejection

Recently, we demonstrated the capacity of allo-specific gene-engineered T lymphocytes as transport vehicle for therapeutic transgenes into allografts. In this study, the influence of viral IL-10 as therapeutic transgene was addressed. Lewis rat T-cell lines specific for DA rat alloantigens were engineered to express vIL-10 by using a retroviral gene expression system. Like T regulatory 1 cells, vIL-10 transgenic T lymphocytes express the phenotype CD4(+)25(+) and secrete, in addition to vIL-10, rat IL-10 and IFN-gamma but no IL-4. First, the capacity of vIL-10 transgenic T-cell lines to modulate alloantigen-specific immune responses was evaluated in vitro. In comparison to control MLR with no transgenic cells or equal numbers of control T(EGFP)-lymphocytes, the proliferation as well as production of IFN-gamma by naive responder cells were significantly diminished. Despite this regulatory capacity in vitro, T(vIL-10)-lymphocytes were not able, either alone or in combination with suboptimal doses of Cyclosporine A, to prolong the survival of either DA rat cardiac or renal allografts in Lewis rat recipients. These data demonstrate that intra-graft IL-10 over-expression is not sufficient to prolong allograft survival in a high-responder strain combination and that the regulatory capacity of T cells in vitro does not predict their in vivo efficiency.     

In vitro generated allospecific cytolytic T lymphocytes injure pancreatic islets

The role of cytotoxic T lymphocytes in the rejection of pancreatic islet allografts remains poorly defined. The present study was designed to assess the ability of in vitro generated cytolytic T lymphocytes to produce allospecific functional and structural damage of mouse pancreatic islets. A mixed lymphocyte-islet coculture model (MLIC) has been developed, in which islets from DBA/2J mice (H-2d) stimulate the generation of allospecific cytolytic T lymphocytes (C57B1/6, H-2b), as measured by lysis of allospecific chromium-labeled tumor targets. Responder C57B1/6 splenocytes sensitized to DBA/2J islets were harvested from the MLIC on Day 5 and cocultured with either freshly isolated DBA/2J or B10.BR (H-2K) islets. Islet injury was determined by assessment of beta cell function after 8 hr (as measured by insulin release in response to a glucose challenge) and islet destruction after 24 hr of coculture with the sensitized splenocytes. Whereas coculture of third party B10.BR islets with MLIC-sensitized C57B1/6 anti-DBA splenocytes had no effect on insulin release or structure, incubation of allospecific DBA/2J islets with these splenocytes resulted in inhibition of insulin release after 8 hr and disintegration of the islets by 24 hr. The depletion of MLIC-sensitized C57B1/6 anti-DBA splenocytes with anti-Lyt2 monoclonal antibody, but not anti-L3T4 monoclonal antibody, prevented the allospecific destruction of fresh islets by the splenocytes in culture. This study suggests that allospecific, cytotoxic T lymphocytes may play an important role in the effector mechanism of pancreatic islet allograft destruction.     

微波烘疗对原发性淋巴水肿免疫细胞的影响

OBJECTIVE: To elucidate the effects of microwave on the immunological cells in primary lymphedema. METHODS: The immunological cells including lymphocytes in the affected limb skin and peripheral blood of 10 patients with primary lower limb lymphedema were analysed using ABC and APAAP immunohistochemical methods before and after microwave baking and bandaging treatment. RESULTS: It is demonstrated that in the peripheral blood of the patients there was an increase of CD8+ T lymphocytes as well as a decrease of CD4+ T lymphocytes and the ratio of CD4/CD8. It was found that there was significant perivascular infiltration of mononuclear cells (most were monocytes and macrophages) in the skin of the affected limb. CONCLUSION: Microwave modulates the systemic immunological imbalance by its heating and complex biological effects on primary lymphedema patients through reversing the ratio of CD4/CD8 to normal level by increasing CD4+ T lymphocytes and decreasing CD8+ T lymphocytes. It can also decrease the perivascular T-lymphocyte infiltration of the affected dermis and enhance the phagocytic capabilities by promoting the proteolytic activities of macrophages, finally resulting in edema resolution.