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Objective To investigate the effect of umbilical cord blood dendritic cells(DCs)induced by gastric cancer antigen combined with cytokine induced killer(CIK)cells in gastric cancer cell lines SGC-7901 in vitro. Methods Mononuclear cells from umbilical cord blood were used to create DCs and CIKs. The cell surface antigen expression of the mature DCs such as CD83,CD86,CD11c and the cell surface antigen of CIKs such as CD3,CD56,CD4,CD8,CD16 were detected using flow cytometry. Sensitized DCs-CIKs,DCs-CIKs,CIKs as effective cells,and SGC-7901 as target cells,the killing activities of these effective cells were tested with LDH release,which the number ratio of cells between effective cells and SGC-7901 cells were 10 :1,20 : 1,40 : 1,respectively. Results The cell surface antigen expressions of the mature DCs,such as CD83 + CD86 + ,CD11c + CD83 + ,CD86 + CD11c + were(75. 4 ± 2. 1)% ,(79. 3 ± 1. 4)% ,(80. 2 ± 2. 6)% , respectively. The mature sensitive-DCs surface antigen expressions,such as CD83 + CD86 + ,CD11c + CD83 + , CD86 + CD11c + ,were(77. 7 ± 1. 5)% ,(82. 6 ± 1. 9)% ,(76. 9 ± 2. 6)% ,respectively. There was no sta-tistical significance about the surface antigen expression between DCs and sensitive-DCs(t = 1. 526,P ﹥ 0. 05;t = 0. 958,P ﹥ 0. 05;t = 1. 049,P ﹥ 0. 05). The CIKs surface antigen expressions,such as CD4 + ,CD8 + , CD3 + CD56 + CD16 + ,were(22. 8 ± 1. 3)% ,(77. 3 ± 1. 8)% ,(24. 5 ± 2. 1)% ,respectively. The results suggested that the killing effect of the three kinds of combination cells on gastric cancer cells was different. The number ratio of cells between sensitive-DCs and SGC-7901 cells were 10 : 1,20 : 1,40 : 1,which the killing activities of sensitive-DCs-CIKs against SGC-7901 were(37. 68 ± 1. 49)% ,(41. 67 ± 0. 90)% ,(42. 71 ± 0. 98)% ,respectively. The killing activity of sensitive-DCs-CIKs was the highest when the ratio of cells between sensitive-DCs and SGC-7901 cells were 40 : 1. The killing activities of DC-CIKs were(36. 77 ± 0. 46)% ,(38. 94 ± 0. 95)% ,(41. 15 ± 0. 89)% ,respectively. The killing activities of CIKs were(34. 74 ± 1. 01)% ,(37. 76 ± 0. 43)% ,(39. 65 ± 0. 79)% ,respectively. There were statistically significant differences among the three groups(F = 5. 92,P ﹤ 0. 05;F = 19. 13,P ﹤ 0. 05;F = 8. 88,P ﹤ 0. 05). Conclusion The tumor killing activity of CIK is enhanced obviously by umbilical cord blood DCs which is sensitized by gastric cancer tumor antigen. There is the highest killing activity when the number ratio of cells between sensitive-DC-CIK and SGC-7901 cells is 40 : 1.
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Objective To observe the clinical efficacy of cytokine-induced killer cells (CIK) combined with three dimensional conformal radiotherapy synchronization XELOX regimen in the treatment of local recurrence of rectal neoplasms.Methods A total of 81 patients with local recurrence of rectal neoplasms were divided into treatment group (41 patients) and control group (40 patients) by random digits table method.In treatment group,41 patients received simultaneous three dimensional conformal radiotherapy combined with XELOX chemotherapy,then received CIK treatment.In control group,40 patients received concurrent chemoradiotherapy.The immune function,quality of life (QOL) scores,recent efficacy,survival rate and adverse reactions were analyzed.Results In treatment group,the level of CD3+,CD4+,CD4+/CD8+ and CD16+ CD56+ increased and the level of CD8+ decreased significantly after treatment than those before treatment (0.671 ± 0.012 vs.0.625 ± 0.022,0.378 ± 0.043 vs.0.315 ± 0.035,1.59 ± 1.41 vs.1.02 ± 1.14,0.184 ±0.008 vs.0.121 +0.023,0.237 ±0.030 vs.0.308 ±0.031,P ＜0.05).Compared with those in control group after treatment,the level of CD3+,CD4+,CD4+/CD8+ and CD16+ CD56+ in treatment group increased and the level of CD8+ decreased significantly (P ＜ 0.05).QOL scores of 70.7％ (29/41) patients in treatment group was improved,significantly higher than the 32.5％ (13/40) in conctol group (P =0.001).The response rate(RR) in treatment group was 92.7％(38/41),significantly higher than that in control group [75.0％ (30/40)] (P =0.030).The 1-year survival rate in treatment group and control group were 90.2％(37/41) and 85.0％(34/40),there was no significant difference (P =0.473).The side effect in treatment group and control group showed no significant difference (P＞0.05).Conclusion CIK combined with concurrent chemoradiotherapy in the treatment of local recurrence of rectal neoplasms can improve patients 'immune function,QOL and recent efficacy.
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Objective: To construct cytokine-induced killer (CIK) cell vehicles carrying recombinant adenovirus carrying tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) gene, and preliminarily observe its anti-hepatoma ability. Methods: Lymphocytes were isolated from peripheral blood to culture CIK cells. The phenotypic identification of CIK cells was performed by flow cytometry (FCM). Then, the lentiviral pLenti-hCD40L-E1AB containing CD40L promoter and the recombined adenovirus vector pAd5/35-TRAIL were constructed, respectively. The two viruses were infected into CIK cells by two-step method. After that, the secretory function and proliferative capacity of CIK cells as well as the effect on angiogenesis and the ablility of colony-formation of hepatoma cells were assessed by ELISA, MTT method, Tubule formation assay and soft agarose assay, respectively. Results: The CIK lymphocytes grew vigorously, in which the expressions of CD3, CD56, CD11a and CD226 were positive, while the expressions of CD8 and CD305 were negative. The lentiviral pLenti-hCD40L-E1AB containing active hCD40L promoter and adenovirus E1 gene was successfully constructed, and the recombined adenovirus vector pAd5/35-TRAIL containing human TRAIL gene was also constructed. In CIK cells infected with Ad5/F35-TRAIL and pLenti-hCD40L-E1AB, the expression level of interferon-? was significanly increased (P < 0.05), and the angiogenesis and the colony-formation rate of hepatoma cells were inhibited, but the proliferative capacity of CIK cells was less affected. Conclusion: CIK cell vehicles carrying adenovirus and expressing TRAIL gene are successfully constructed. The growth inhibition of hepatoma cells may be induced by CIK cells. Copyright © 2013 by TUMOR.
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ObjectiveTo investigate the effect of the immunotherapy of CIK cell, LAK cell and PBLS cell mediated by anti-EGFR/anti-CD3 bispecific antibody (BsAb) respectively on the mice borne human gastric cancer and provide experimental evidence for therapeutic strategy in treating gastric cancer. Methods The mAbs of anti-CD3 and anti-EGFR were cross-linked to prepare the BsAb by chemical synthesis. The experimental therapy on the mice borne SGC7901 human gastric cancer was performed,and then the comparisons of the curative activity among the CIK group, LAK group and PBLS group were conducted in vivo. ResultsThe mean tumor reduction rate of the administration of CIK cells directed by anti-EGFR/anti-CD3 BsAb was(64.9±7.7)％ and higher than those of LAK cells or PBLS targeted by anti-EGFR/anti-CD3 BsAb [(43.5±8.2) ％ and (39.7±6.5) ％] (P ＜ 0.05).The mean tumor weight of the administration of CIK cells directed by anti-EGFR/anti-CD3 BsAb was (473.9±37.7) mg at the end of therapy and was lower than those of LAK cells or PBLS targeted by anti-EGFR/anti-CD3 BsAb [(764.6±88.3) mg and (829.1±104.4) mg](P ＜ 0.05). ConclusionThe CIK cell mediated by anti-EGFR/anti-CD3 BsAb could have better curative effect than other effector cells on gastric cancer in vivo.
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Having an understanding of the properties of cytokines is essential for the immunologist, the researcher and the medical practitioner who need to understand immunologic diseases and immunological therapeutic approaches. Cytokines are redundant in their actions on target cells and promiscuous in their receptor reactions. (ED note: That is some cool use of English!) Moreover, many cells concomitantly produce several cytokines that have overlapping actions. Here this review provides conceptual framework to understand the intriguing aspects of the cytokine system.
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Cytokines , Immune System Diseases , InterleukinsABSTRACT
Objective:To investigate whether the susceptibility of human colon adenocarcinoma cell SW1116 to lymphokine-activated killer cell (LAK)-mediated lysis could be enhanced by low concentration of sodium butyrate, and the possible involvement of intercellular surface adhesion molecule-1 (ICAM-1) and carcinoembryonic antigen (CEA). Methods:Standard MTT assay was used to evaluate the cytotoxic activity of LAK cells to SW1116 cells, flow cytometric and immunofluorescent techniques were used to determine the expression of ICAM-1 and CEA on tumor cells. Results:Treatment of SW1116 with sodium butyrate leads to an increased resistance to LAK-mediated lysis, accompanied by downregulation of ICAM-1 expression and upregulation of CEA expression. Conclusion: Sodium butyrate inhibits rather than enhance LAK activity against SW1116, probably by changing the expression of ICAM-1 and CEA on target cells, which impair the adherence of effector on target cells.
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Objective To investigate the feasibility of lymphokine activated killer(LAK) cells induced from cord blood used as adoptive cellular immunotherapy for human cancer.Methods Mononuclear cells were separated from umbilical blood(UBMC) by Ficoll,and stimulated by IL-2.The phenotypes(CD3/ CD4/ CD8) of the mononuclear cells were assayed by Flow cytometry,and their lethal activity on K562 or SKOV6 assayed by MTT colometric.The peripheral blood mononuclear cells were used as the control.Results The in vitro anti-tumor effect of LAK from cord blood was significant.Conclusion LAK from cord blood can be a source of adoptive cellular immunotherapy in the treatment of human cancer.
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It is well known that the murine T helper cell clones are divided by their lymphokine secretory activities. One is the Th-1 cell, producing IL-2 and IFN after stimulation and the other is the Th-2 cell, producing the IL-4 and IL-5. This study was undertaken to evaluate the immunomodulatory properties of the bacterial lipopolysaccharide(LPS) on the lymphokine production in vivo and in vitro. The results were as follows: There were no effects on the lymphokine secretion by the in vitro treatment of the LPS. The in vivo treatment of the LPS decreases the capability of the production of IL-2 and IFN , whereas it increases the capability of IL-4 production. The altered capacity of the lymphokine production was recovered about 2 weeks after the treatment of the LPS. There were no differences on the lymphokine production between E-coli LPS and salmonella LPS. The capacity of the lymphokine production was the same in the treatment of a non-heated LPS or heated-LPS. The lymphokine production of the mice which were desensitized by the long term treatment of the LPS was not different from the control mice. The in vitro treatment of RU486 can block the alterations of the lymphokine production after the treatment of the LPS. In summary, one can tell that the LPS increases the secretion of the IL-4 through the endogenous secretion of the glucocorticoids.
Subject(s)
Animals , Mice , Clone Cells , Glucocorticoids , Interleukin-2 , Interleukin-4 , Interleukin-5 , Mifepristone , Salmonella , T-Lymphocytes , T-Lymphocytes, Helper-InducerABSTRACT
The peripheral blood lymphocytes (PBLs) from 111 cancer patients were isolated and cultured respectively for 23 - 27 days in the medium mainly conditioned by IL-2 and PHA. With ~(125) I-UdR release method, sampling in random way, we examed the cytotoxicities of PBLs and lymphokine-activated-killer (LAK) cells in different culture periods in vitro. The statistic analysis on sufficient data gave the following results: 1. The cytotoxicity against K562 cells increased from 34.78 ?25% of the PBLs to 68.04 ?17.3% of the cells cultured for 8-13 days, afterward, kept about 70% to 23 - 27 days. The constitutional proportion patterns showed that the freshly isolated samples dispersed at a wide range of cytotoxicities (10 - 90%), and that most of the cultured samples ( ~ 85%) concentrated on the range of higher cytotoxicities (50 ~ 95% ) after 8-13 days. 2. The cytotoxicity against Raji cells rose from 8.9 ?8% of the fresh PBLs to 42.1 ?22% of the LAK cells at 8 - 13 days, and maintained about 35% in the following periods. The constitutional proportion patterns of the cytotoxicity against Raji illustrated that all the fresh PBL samples were below 25% of cytotoxicity, and that during the culture, one part of the samples ( ~ 30%) acquired the higher cytotoxicities (50 -90% ), but the other part of the samples ( - 40%) remained at lower cytotoxicities (below 35% ) . The mechanisms behind these phenomena are worth further investigating.
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In order to study the enhancement of immune functions and autologous tumor-killing (ATX) activity by kappa-selenocarrageenan (KSC) in mice bearing sarcoma 180, the effects of KSC and/or Cyclophosphamide (Cy) on natural killer(NK) activity, lymphokine activated killer(LAK) activity, the production of interleukin-2 (IL-2), ATK activity and the growth of sarcoma 180 (S180) were observed. The results showed that KSC promoted NK activity, LAK activity and ATK activity in vivo, increased IL-2 production at 40mg/kg ?d x 9d. It also enhanced the antitumor action of Cy (20mg/kg?d x 9d) and offsetted the inhibition of Cy on immunocompetent cells. The ATK activity in splenocytes of SI80-bearing mice could be induced and augmented by recombinant interleukin-2 (rIL-2) in vitro. In conclution, KSC had a up-regulating effect on immune functions and ATK activity in tumor-bearing mice, therefore, can be used as a biological response modifier (BRM) in cancer biotherapy.
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Using ovarian cancer cell lines, we studied relationship between C-erbB2 expression and lysis activity of lymphokine-activated killer cells (LAK) and in vitro regulation by ?-interferon (IFN-?). The results showed that C-erbB2 overexpression was associated with resistance to lysis activity of LAK in ovarian cancer; IFN-? could reduce the overexpression of C-erbB2 and increase lysis activity of LAK on ovarian cancer cells in which C-erbB2 were overexpressed.
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In this study, we demonstrated that immobilized fibronectin (FN) enhanced LAK activity, and that the enhanced LAK activity was completely abrogated by an anti-VLA-5 monoclonal antibody and RGD peptide. Fresh -spleen cells expressed VLA-4, VLA-6 and vitronectin receptor, whereas VLA-5 was expressed only on the spleen cells activated with IL-2. LAK cells showed increased adhesion to immobilized FN compared with that to control BSA, and the increased adhesion of LAK cells to immobilized FN was inhibited by anti-VLA-5 monoclonal antibody. Conjugate-formation assay showed that the LAK cells cultured on immobilized FN bound to target cells more efficiently than the control LAK cells, and that anti-LFA-1 monoclonal antibody inhibited the LAK-target cell binding. Immobilized type IV collagen and laminin, as well as FN, enhanced LAK activity. All these results suggest that the interaction of inte-grins expressed on LAK cells with extracellular matrix proteins act as co-stimulator for the enhancement of LAK activity , and that anchorage is necessary for full activation of LAK cells.
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Under stimulation of PHA,PHA + TPA,or PHA + PTS the time ke-netics of DNA,RNA,protein synthesis and lymphokine secretion by human lymph node cell expressed different responses. The PTS did not increase the DNA and RNA synthesis, but increased the protein synthesis ( 60% ) . TPA increased the RNA synthesis ( 18% ) , therefor increased the protein synthesis ( 40% ) . Both of TPA and PTS increased the production of 5 kinds of lymphokines (IL-1 , IL-2 , IL-3, BCGF, TFNr). We infer that PTS and TPA increase the lymphokine production through different gene regulation. TPA Promotes the translation of lymphokine mRNA。
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The survival of implanted tumor cells in mice which had been treated with interferon in combination with either adriamycin or vincristine was evaluated. While the majority of tumor cells implanted into normal mice failed to survive (52.1 to 63.5%), most of those implanted into mice which had been pretreated with either adriamycin or vincristine survived. If the mice were secondarily treated with interferon, the ability of adriamycin or vincristine to inhibit the survival of implanted tumor cells was restored within 24 hours. Restoration of tumoricidal activity by interferon treatment was more evident in the adriamycin pretreated mice. Peritoneal macrophages isolated from mice pretreated with both interferon and adriamycin had an increased tumoricidal activity, when compared with those isolated from mice treated with adriamycin alone. This interferon dependent enhancement of tumoricidal activity was comparable with that obtained by treating mice with lymphokines a product of Con A treated lymphocytes isolated from BCG treated mice. These results suggested that both adriamycin and vincristine may damage the macrophages required for the natural host defense mechanism and allow the implanted tumor cells to survive. Interferon may, however, protect the macrophages from drug induced damage.
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Mice , Animals , Doxorubicin/therapeutic use , Interferon Type I/therapeutic use , Macrophages/immunology , Mice, Inbred ICR , Neoplasms, Experimental/therapy , Vincristine/therapeutic useABSTRACT
TIL were isolated from the resected tumor mass,and lymphocytes were separated from pe-ripheral blood of 12 patients with osteosarcoma.In vitro antitumor activity and spesificity of rIL—2 activated TIL and LAK cells were determined by 4 hour ~(51)Cr releasing assay.Phenotypeanalysis of TIL were carried out in different intervals of incubation.The results revealed thatduring the incubation period from Day 15—Day 20,the difference between average cytolytic ac-tivities of TIL and LAK ceils with K562 and LiBr cells as target cells(E:T ratio 25:1)were in-significant,while that from 7 of those patients with autotumor cells as target cells were signifi-cant.The phenotype analysis showed that the percentage of CD3~+ cells remained constant,whilethat of CD4~+ cells tends to increase,and that of CD8~+cells tends to decrease during the whole in-cubation period.
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PHA,PMA or OKT3 McAb alone can not induce the proliferative responses of purified resting T cells,which enables us to comparise the effects of exogenous IL—2,IL—4 and IL—6 on the activation and proliferation of T cell.PHA can induce purifeid resting T cells responding to exgenous IL—2,IL—4 or Il—6 and anti—Tac(p55)McAb only blocked the proliferative response of PHA—stimulated T cells responding to IL—2.PMA can induce purified resting T cells responding to exogenous IL—2 or IL—4 but not Il—6.OKT3 McAb only induced the purified resting T cells responding to exogenous IL—2.Cyclsporin A,an immunosuppressant(CsA),inhibited the proliferative responses of PHA—stimulated T cells responding to exogenous IL—2,IL—4 or IL—6,but in the kinetic study of inhibitory affect of CsA,the results were completely different.Our data suggest that IL—2,IL—4 and IL—6 activate T cell by different pathway.
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We previously demonstrated the cytotoxicity of human LAK cells to solid tumors in vitro. The work we reported here investigated the mechanisms involved in killing of solid tumor cells by LAK cells. It was shown that the cytotoxicity of LAK cells was mediated by some factors secreted by LAK cells and effects of direct contact with targets. Following lysis, the nucli of targets were destroyed and fragments of DNA were released into the medium. The cytotoxicity of LAK cells, under some circumtances, was of a positive correlation to the proliferation of themselves.
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We have investigated, in both cases of normal individuals and tumor- bearing patients, the cytotoxic activity of lymphokine- activated killer (LAK) cells to cultured cell lines or fresh solid tumor cells with a 4- hour chromium 51 (~(51)Cr) - release assay. The LAK cells were generated by incubating peripheral blood mononuclear (PBM) cells in vitro with recombinant interleukin 2 (rIL- 2)or partially purified interleukin2(PPIL- 2). Our results indicated that these two sources of L \K cells were cytotoxic to either K562, NK- sensitive or Daudi, NK resistant tumor cell lines. These two kinds of LAK cells could also kill a broad rang of fresh solid tumor cells, suggesting that these cells be indeed LAK cells. Furthermore, the results demonstreated that some fresh solid tumor cells were resistant to LAK cell killing to some extent as they showed different sensitivities to LAK cytotoxicity.
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It was demonstrated that TILs exhibited higher cytotoxicity to autologous tumor cells than that of LAK cells grown under identical conditions, and TILs had target cell specificity.The cytotoxicity of TILs to other tumor cells was lower than that of LAK cells, and TILs had no cytotoxicity to ConA-induced lymphoblast cells but LAK cells did have some cytotoxicity to the normal cells. The cytotoxicity of TILs to B 16 melanoma cells was reduced by monoclonal antimelanoma antibodies (M2590 and M562) which can block the activity of antimelanoma CTL, suggesting TILs contained high level of CTL activity. TILs were found to be more effective in their therapeutic potency in vivo on a percell basis than were LAK cells. These results indicate that TILs are more suitable to the adoptive immunotherapy of turn or as effector cells than LAK cells.
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Leukoregulin is an immunologic hormone whose anticancer action includethe inhibition of cellular proliferation and cytolysis of tumor cells eitherdirectly or indirectly by enhancing the tumor cell's susceptibility to dest-ruction mediated by natural killer cells.The optimal condition for the ind-uction of leukoregulin Were studied by using orthogonal experimental design.The optimal concentration is 1x10~7/ml for human spleen cells;3.1?g/mlfor phytohemagglutinin(PHA);12.5ng/ml for 12-0-tetradecanoyl-phorbol-13-acetate;15% for boyin serum.Leukoregulin activity prepared by 70 hincubation is maximum,the experiment also confirmed that the inducingleukoregulin effect of PHA-PⅢis significantly high.