mixed lymphocyte culture test

Summary

Summary: Measure of histocompatibility at the HL-A locus. Peripheral blood lymphocytes from two individuals are mixed together in tissue culture for several days. Lymphocytes from incompatible individuals will stimulate each other to proliferate significantly (measured by tritiated thymidine uptake) whereas those from compatible individuals will not. In the one-way MLC test, the lymphocytes from one of the individuals are inactivated (usually by treatment with MITOMYCIN or radiation) thereby allowing only the untreated remaining population of cells to proliferate in response to foreign histocompatibility antigens.

Top Publications

  1. Hoogduijn M, Crop M, Peeters A, van Osch G, Balk A, Ijzermans J, et al. Human heart, spleen, and perirenal fat-derived mesenchymal stem cells have immunomodulatory capacities. Stem Cells Dev. 2007;16:597-604 pubmed
    ..The results of this study demonstrate that MSCs from various tissues have similar immunophenotypes, in vitro immunosuppressive properties, and differentiation potential. ..
  2. Tsukahara T, Kawaguchi S, Torigoe T, Kimura S, Murase M, Ichimiya S, et al. Prognostic impact and immunogenicity of a novel osteosarcoma antigen, papillomavirus binding factor, in patients with osteosarcoma. Cancer Sci. 2008;99:368-75 pubmed publisher
    ..These findings suggested prognostic significance and immunodominancy of PBF in patients with osteosarcoma. PBF is the candidate target for immunotherapy in patients with osteosarcoma. ..
  3. Cho P, Messina D, Hirsh E, Chi N, Goldman S, Lo D, et al. Immunogenicity of umbilical cord tissue derived cells. Blood. 2008;111:430-8 pubmed
  4. AgauguƩ S, Perrin Cocon L, Andre P, Lotteau V. Hepatitis C lipo-Viro-particle from chronically infected patients interferes with TLR4 signaling in dendritic cell. PLoS ONE. 2007;2:e330 pubmed
    ..By acting on TLR4 pathway with LVP, HCV may thus exploit a natural protective mechanism of the liver and the intestine normally used to control inflammation and immunity to commensal microorganisms. ..
  5. Hipp M, Hilf N, Walter S, Werth D, Brauer K, Radsak M, et al. Sorafenib, but not sunitinib, affects function of dendritic cells and induction of primary immune responses. Blood. 2008;111:5610-20 pubmed publisher
    ..These results indicate that sunitinib, but not sorafenib, is suitable for combination with immunotherapeutic approaches for treatment of cancer patients. ..
  6. Onai N, Obata Onai A, Schmid M, Ohteki T, Jarrossay D, Manz M. Identification of clonogenic common Flt3+M-CSFR+ plasmacytoid and conventional dendritic cell progenitors in mouse bone marrow. Nat Immunol. 2007;8:1207-16 pubmed
    ..These clonogenic common DC progenitors thus define a cytokine-regulated DC developmental pathway that ensures the supply of various DC populations. ..
  7. Weiss M, Anderson C, Medicetty S, Seshareddy K, Weiss R, VanderWerff I, et al. Immune properties of human umbilical cord Wharton's jelly-derived cells. Stem Cells. 2008;26:2865-74 pubmed publisher
    ..The results suggest that human UCMS will be tolerated in allogeneic transplantation. Disclosure of potential conflicts of interest is found at the end of this article. ..
  8. Ryan J, Barry F, Murphy J, Mahon B. Interferon-gamma does not break, but promotes the immunosuppressive capacity of adult human mesenchymal stem cells. Clin Exp Immunol. 2007;149:353-63 pubmed
  9. Magatti M, De Munari S, Vertua E, Gibelli L, Wengler G, Parolini O. Human amnion mesenchyme harbors cells with allogeneic T-cell suppression and stimulation capabilities. Stem Cells. 2008;26:182-92 pubmed

More Information

Publications62

  1. Waggoner S, Hall C, Hahn Y. HCV core protein interaction with gC1q receptor inhibits Th1 differentiation of CD4+ T cells via suppression of dendritic cell IL-12 production. J Leukoc Biol. 2007;82:1407-19 pubmed
    ..Importantly, that addition of IL-12 rescued IFN-gamma production and Th1 differentiation by CD4(+) T cells. Therefore, engagement of gC1qR on DCs by HCV core limits the induction of Th1 responses and may contribute to viral persistence. ..
  2. English K, Barry F, Field Corbett C, Mahon B. IFN-gamma and TNF-alpha differentially regulate immunomodulation by murine mesenchymal stem cells. Immunol Lett. 2007;110:91-100 pubmed
  3. Popov I, Li M, Zheng X, San H, Zhang X, Ichim T, et al. Preventing autoimmune arthritis using antigen-specific immature dendritic cells: a novel tolerogenic vaccine. Arthritis Res Ther. 2006;8:R141 pubmed
    ..In conclusion, we demonstrate that in vitro-generated antigen-specific immature DC may have important potential as a tolerogenic vaccine for the treatment of autoimmune arthritis. ..
  4. Meyer T, Zehnter I, Hofmann B, Zaisserer J, Burkhart J, Rapp S, et al. Filter Buffy Coats (FBC): a source of peripheral blood leukocytes recovered from leukocyte depletion filters. J Immunol Methods. 2005;307:150-66 pubmed
    ..In summary, Filter Buffy Coats are a valuable and convenient source of different peripheral leukocyte populations and can replace standard buffy coat preparations for research applications. ..
  5. Eliopoulos N, Stagg J, Lejeune L, Pommey S, Galipeau J. Allogeneic marrow stromal cells are immune rejected by MHC class I- and class II-mismatched recipient mice. Blood. 2005;106:4057-65 pubmed
    ..These results strongly suggest that MSCs are not intrinsically immunoprivileged and cannot serve as a "universal donor" in immunocompetent MHC-mismatched recipients. ..
  6. Averill L, Lee W, Karandikar N. Differential dysfunction in dendritic cell subsets during chronic HCV infection. Clin Immunol. 2007;123:40-9 pubmed
    ..Our studies demonstrate distinct and contrasting dysfunctional features in DC subsets and underscore the importance of evaluating APC subpopulations separately. ..
  7. Tripathi S, Maier K, Bruch D, Kittur D. Effect of 6-gingerol on pro-inflammatory cytokine production and costimulatory molecule expression in murine peritoneal macrophages. J Surg Res. 2007;138:209-13 pubmed
  8. Kalthoff F, Winiski A, Fichtinger P, Schwendinger B, Wang S, Weishaeupl C, et al. Differential inhibition of primary versus preactivated T cells by pimecrolimus but not by tacrolimus in vitro. Int Arch Allergy Immunol. 2007;142:255-64 pubmed
    ..These data indicate that pimecrolimus has a selectivity for antigen-primed memory T cells not seen with tacrolimus. ..
  9. Tripathi S, Bruch D, Kittur D. Ginger extract inhibits LPS induced macrophage activation and function. BMC Complement Altern Med. 2008;8:1 pubmed publisher
    ..A significant decrease in IFN-gamma and IL-2 production by T cells in response to allostimulation was also observed. In conclusion ginger extract inhibits macrophage activation and APC function and indirectly inhibits T cell activation. ..
  10. Cho H, Choi E, Lee S, Jung K, Seo S, Choi I, et al. Programmed death-1 receptor negatively regulates LPS-mediated IL-12 production and differentiation of murine macrophage RAW264.7 cells. Immunol Lett. 2009;127:39-47 pubmed publisher
    ..7 cells upon PD-1 engagement. Taken together, our results reveal a novel function of macrophage PD-1 in the negative regulation of IL-12 synthesis and differentiation into dendritic cell-like cells. ..
  11. Wang Y, Chen X, Armstrong M, Li G. Survival of bone marrow-derived mesenchymal stem cells in a xenotransplantation model. J Orthop Res. 2007;25:926-32 pubmed
    ..The data suggested that rat MSCs survived the 11 weeks of xenotransplantation in the MF1 mice, but the increased host immune sensitization led to the impaired in vivo osteogenesis potential of MSCs. ..
  12. Glouchkova L, Ackermann B, Zibert A, Meisel R, Siepermann M, Janka Schaub G, et al. The CD70/CD27 pathway is critical for stimulation of an effective cytotoxic T cell response against B cell precursor acute lymphoblastic leukemia. J Immunol. 2009;182:718-25 pubmed
    ..Modulation of the CD70/CD27 pathway may thus represent a novel therapeutic approach for augmenting magnitude and quality of the antileukemic response in B cell precursor acute lymphoblastic leukemia. ..
  13. Schuster C, Vaculik C, Fiala C, Meindl S, Brandt O, Imhof M, et al. HLA-DR+ leukocytes acquire CD1 antigens in embryonic and fetal human skin and contain functional antigen-presenting cells. J Exp Med. 2009;206:169-81 pubmed publisher
    ..Collectively, our data provide insight into skin DC biology and the mechanisms through which skin DCs presumably populate the skin during development. ..
  14. Micklethwaite K, Garvin F, Kariotis M, Yee L, Hansen A, Antonenas V, et al. Clinical-scale elutriation as a means of enriching antigen-presenting cells and manipulating alloreactivity. Cytotherapy. 2009;11:218-28 pubmed publisher
    ..In addition to providing enrichment of monocytes for the generation of DC, the Elutra enriches cell subsets that may be incorporated into and enhance existing immunotherapy and stem cell transplantation protocols. ..
  15. Yu H, Shen G, Wei F. Effect of cryopreservation on the immunogenicity of osteoblasts. Transplant Proc. 2007;39:3030-1 pubmed
    ..Fluid nitrogen cryopreservation is an ideal method to reduce the immunogenicity in allograft. ..
  16. Tran C, Burton L, Russom D, Wagner J, Jensen M, Forman S, et al. Manufacturing of large numbers of patient-specific T cells for adoptive immunotherapy: an approach to improving product safety, composition, and production capacity. J Immunother. 2007;30:644-54 pubmed
    ..These new methods allow for more efficient production multiple patient-specific products by satisfying the basic tenants of safety and efficacy required for early phase clinical trials of cell products. ..
  17. Xiang J, Gu X, Qian S, Chen Z. Graded function of CD80 and CD86 in initiation of T-cell immune response and cardiac allograft survival. Transpl Int. 2008;21:163-8 pubmed
    ..In this study, graded stimulatory function of CD80 and CD86, stronger inhibition of proliferation, and stronger prolongation of transplant survival were observed when CD80 and CD86 were blocked simultaneously. ..
  18. Stroopinsky D, Avivi I, Rowe J, Avigan D, Katz T. Allogeneic induced human FOXP3(+)IFN-gamma(+) T cells exhibit selective suppressive capacity. Eur J Immunol. 2009;39:2703-15 pubmed publisher
    ..Such unique properties suggest that these cells are potentially ideal for the use as post-transplant GVH disease prophylaxis. ..
  19. de Bold M, Etchepare A, Martinuk A, De Bold A. Cardiac hormones ANF and BNP modulate proliferation in the unidirectional mixed lymphocyte reaction. J Heart Lung Transplant. 2010;29:323-6 pubmed publisher
    ..The modulation of the MLR by NP suggests a possible clinical use of these peptides in transplantation immunity. ..
  20. Oikawa A, Ito K, Seguchi H, Okabe M, Migishima F, Eshima K, et al. Development of immunocompetent lymphocytes in vivo from murine umbilical cord blood cells. Transplantation. 2007;84:23-30 pubmed
    ..Collectively the data indicated that T and B lymphocytes derived from UCBC-HSC are fully competent in immunological terms. ..
  21. Xiang M, Zou X, Zhang C, Zhao Z, Xu J. Insulin administration confers diabetes-preventive properties to NOD mice derived dendritic cells. Immunopharmacol Immunotoxicol. 2007;29:451-64 pubmed
    ..These results suggest that a subtype of DC generated by insulin subcutaneous treated NOD mice confers potential protection from diabetes through polarizing the immune response towards a Th2 regulatory pathway. ..
  22. Lei C, Ou Y, Yang Z, Tang X. [The infection of dendritic cells by recombinant adenoviral vector carrying HBsAg-HSP70 chimeric gene and its biological characteristics observations]. Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2009;23:29-31 pubmed
    ..MLR showed that there are not a statitic difference of stimulated index (SI) between two groups. Results indicated that Ad-HSP70-HBsAg can effectively infected DCs without affecting its biological characteristics. ..
  23. Sabbagh F, Lecerf F, Hulin A, Bac P, German Fattal M. Effect of hypomagnesemia on allogeneic activation in mice. Transpl Immunol. 2008;20:83-7 pubmed publisher
    ..We hypothesize that the remaining activity is sufficient for IL-2 gene normal activation. Alternatively, Mg-deficiency may trigger other signaling pathways leading to IL-2 production. ..
  24. Wang W, Carper K, Malone F, Latchman Y, Perkins J, Fu Y, et al. PD-L1/PD-1 signal deficiency promotes allogeneic immune responses and accelerates heart allograft rejection. Transplantation. 2008;86:836-44 pubmed publisher
    ..Absence of PD-L1 input triggers a stimulatory signal to effector T cells and APCs, accelerating heart allograft rejection. Engagement of the PD-L1 signal on T cells or APCs may be necessary to induce transplant tolerance. ..
  25. Obermajer N, Repnik U, Jevnikar Z, Turk B, Kreft M, Kos J. Cysteine protease cathepsin X modulates immune response via activation of beta2 integrins. Immunology. 2008;124:76-88 pubmed publisher
    ..We demonstrate that cathepsin X is involved in the regulation of two beta2 integrin receptors, LFA-1 and Mac-1, which exhibit opposing roles in lymphocyte activation. ..
  26. Wenink M, Santegoets K, Broen J, van Bon L, Abdollahi Roodsaz S, Popa C, et al. TLR2 promotes Th2/Th17 responses via TLR4 and TLR7/8 by abrogating the type I IFN amplification loop. J Immunol. 2009;183:6960-70 pubmed publisher
    ..This shift in T cell responses was inherent to the inability of TLR2-stimulated DCs to produce IL-12p70 and was dependent on the production of IL-1 and IL-6. ..
  27. Hutchinson J, Roelen D, Riquelme P, Brem Exner B, Witzke O, Philipp T, et al. Preoperative treatment of a presensitized kidney transplant recipient with donor-derived transplant acceptance-inducing cells. Transpl Int. 2008;21:808-13 pubmed publisher
    ..The case of patient FR provides further evidence of the safety of pre-transplantation treatment with TAICs. ..
  28. Kim S, Kim H, Kim H, Lee K, Kim H. Generation of functionally mature dendritic cells from elutriated monocytes using polyinosinic : polycytidylic acid and soluble CD40 ligand for clinical application. Clin Exp Immunol. 2008;154:365-74 pubmed publisher
    ..Thus, the protocol based on the combination of poly(I:C) and CD40L is more effective for the induction of clinical-grade DCs from elutriated monocytes than the standard cytokine cocktail. ..
  29. Zhang D, Sanchez Fueyo A, Kawamoto K, Alexopoulos S, Zhang W, Zheng X. Th1 to Th2 immune deviation facilitates, but does not cause, islet allograft tolerance in mice. Cytokine. 2010;51:311-9 pubmed publisher
    ..Thus, anti-IL-12 and low-dose IL-4/Fc facilitate, but do not cause, islet allograft tolerance in mice by increasing the immunosuppressive potency of CD4(+)CD25(+) regulatory T cells. ..
  30. Methe H, Hess S, Edelman E. Endothelial cell-matrix interactions determine maturation of dendritic cells. Eur J Immunol. 2007;37:1773-84 pubmed
    ..We now demonstrate for the first time that interactions of EC with an underlying 3D matrix affect the ability of EC to drive DC maturation. ..
  31. Xu Y, Huang H. [Effects on activity of mouse NK cells by inhibitory Ly49 receptors blockade]. Zhejiang Da Xue Xue Bao Yi Xue Ban. 2009;38:130-5 pubmed
    ..014), and statistical difference was observed only between antibody groups of 0 microg/ml and 60 microg/ml (P<0.05). Blocking the inhibitory Ly49 receptor on NK cells with monoclonal antibody augments the cytotoxicity to target cells. ..
  32. Zheng P, Yang Y, Li S, Li J, Gong W, Quan Z. A given number of effector T cells can only destroy a limited number of target cells in graft rejection. Transpl Immunol. 2010;23:111-6 pubmed publisher
  33. Wada N, Menicanin D, Shi S, Bartold P, Gronthos S. Immunomodulatory properties of human periodontal ligament stem cells. J Cell Physiol. 2009;219:667-76 pubmed publisher
    ..These results suggest that PDLSCs, BMSSCs, DPSCs, and GFs possess immunosuppressive properties mediated, in part, by soluble factors, produced by activated PBMNCs. J. Cell. Physiol. 219: 667-676, 2009. (c) 2009 Wiley-Liss, Inc. ..
  34. Hendrikx T, van Gurp E, Mol W, Schoordijk W, Sewgobind V, Ijzermans J, et al. End-stage renal failure and regulatory activities of CD4+CD25bright+FoxP3+ T-cells. Nephrol Dial Transplant. 2009;24:1969-78 pubmed publisher
    ..It now appears that in this setting, regulation by CD4(+)CD25(bright+)FoxP3(+) T-cells is also impaired. ..
  35. Maeda M, Ito Y, Hatanaka T, Hashiguchi S, Torikai M, Nakashima T, et al. Regulation of T cell response by blocking the ICOS signal with the B7RP-1-specific small antibody fragment isolated from human antibody phage library. MAbs. 2009;1:453-61 pubmed
  36. Szabo G, Mandrekar P. Human monocytes, macrophages, and dendritic cells: alcohol treatment methods. Methods Mol Biol. 2008;447:113-24 pubmed publisher
    ..Here, we summarize the different effects of acute and chronic alcohol on monocyte, macrophage, and dendritic cell functions in humans and describe methods for separation and functional evaluation of these cell types. ..
  37. Gredler V, Ebner S, Schanda K, Forstner M, Berger T, Romani N, et al. Impact of human myelin on the maturation and function of human monocyte-derived dendritic cells. Clin Immunol. 2010;134:296-304 pubmed publisher
    ..We hypothesize that myelin influences DC activation and plays a pivotal role in balancing immunity and tolerance. ..
  38. Elluru S, Duong Van Huyen J, Delignat S, Kazatchkine M, Friboulet A, Kaveri S, et al. Induction of maturation and activation of human dendritic cells: a mechanism underlying the beneficial effect of Viscum album as complimentary therapy in cancer. BMC Cancer. 2008;8:161 pubmed publisher
    ..In the present study, we examine the hypothesis that the VA preparations induce activation of human DC that facilitates effective tumor regression...
  39. Auerbach M, Shimoda N, Amano H, Rosenblum J, Kish D, Farber J, et al. Monokine induced by interferon-gamma (MIG/CXCL9) is derived from both donor and recipient sources during rejection of class II major histocompatibility complex disparate skin allografts. Am J Pathol. 2009;174:2172-81 pubmed publisher
    ..These results therefore demonstrate the synergy of donor- and recipient-derived Mig in promoting T-cell infiltration into allografts. ..
  40. Truong D, Cornet A, Wieƫrs G, Robert A, Reding R, Latinne D. Pre- and post-transplant monitoring of granzyme B enzyme-linked immunosorbent spot assay in pediatric liver recipients. Transpl Immunol. 2008;19:215-9 pubmed publisher
    ..However, a kinetic study of GrB ELISPOT could be helpful to predict or confirm early rejection in the small group of liver allograft recipients analyzed in this study. ..
  41. Wan C, Cheng R, Wang H, Liu T. Immunomodulatory effects of mesenchymal stem cells derived from adipose tissues in a rat orthotopic liver transplantation model. Hepatobiliary Pancreat Dis Int. 2008;7:29-33 pubmed
    ..05). Adipose-derived MSCs clearly inhibit recipient-derived T lymphocyte proliferation in MLC and significantly alleviate acute rejection following orthotopic liver transplantation in rats. ..
  42. Takahashi H, Iwagaki H, Hamano R, Kanke T, Liu K, Sadamori H, et al. Effect of adenosine receptor subtypes stimulation on mixed lymphocyte reaction. Eur J Pharmacol. 2007;564:204-10 pubmed
    ..These results suggest that adenosine inhibits the immune responses during mixed lymphocyte reaction via adenosine A(2A) receptor. ..
  43. Matsushima H, Tanaka H, Mizumoto N, Takashima A. Identification of crassin acetate as a new immunosuppressant triggering heme oxygenase-1 expression in dendritic cells. Blood. 2009;114:64-73 pubmed publisher
  44. Jochum C, Beste M, Stone D, Graves S, Storb R. Development and in vitro characterization of canine CD40-Ig. Vet Immunol Immunopathol. 2008;123:260-5 pubmed publisher
    ..Canine CD40-Ig is more immunosuppressive than the anti-human CD154 antibody 5c8 in canine mixed leukocyte reactions and may be more effective in vivo in a model of marrow transplantation. ..
  45. Ohashi K, Takahashi H, Mori S, Liu K, Wake H, Sadamori H, et al. Advanced glycation end products enhance monocyte activation during human mixed lymphocyte reaction. Clin Immunol. 2010;134:345-53 pubmed publisher
    ..AGE-2 and AGE-3 up-regulated the expression of RAGE on monocytes; therefore, the AGEs may activate monocytes, leading to the up-regulation of adhesion molecule expression, cytokine production and T-cell proliferation. ..
  46. Park Y, Koo S, Kim Y, Kim H, Joe I, Park C, et al. Effect of in vitroexpanded CD4(+)CD25(+)Foxp3(+) regulatory T cell therapy combined with lymphodepletion in murine skin allotransplantation. Clin Immunol. 2010;135:43-54 pubmed publisher
    ..We conclude that Treg cell therapy combined with lymphodepletion is of practical benefit for the control of allograft rejection, but requires supplementary measures to promote immune tolerance. ..
  47. Gillet Hladky S, Duperrier K, Picandet S, Mathias V, de Carvalho M, Bernaud J, et al. Induction of high expression of CCR7 and high production of IL-12 in human monocyte-derived dendritic cells by a new bacterial component: LCOS 1013. Int Immunopharmacol. 2008;8:1151-63 pubmed publisher
    ..Moreover, we found that LCOS 1013 activates DCs through TLR2. LCOS 1013 represents an attractive therapeutic maturation agent of DCs allowing the production of Mo-DCs with high capacity to migrate and to induced Th1 immune responses. ..
  48. Vucevic D, Melliou E, Vasilijic S, Gasic S, Ivanovski P, Chinou I, et al. Fatty acids isolated from royal jelly modulate dendritic cell-mediated immune response in vitro. Int Immunopharmacol. 2007;7:1211-20 pubmed
    ..In conclusion, our results showed the immunomodulatory activity of RJ fatty acids and suggest that DC are a significant target of their action. ..
  49. Huang Y, Chen X, Dikov M, Novitskiy S, Mosse C, Yang L, et al. Distinct roles of VEGFR-1 and VEGFR-2 in the aberrant hematopoiesis associated with elevated levels of VEGF. Blood. 2007;110:624-31 pubmed
    ..Our findings suggest that selective blockade of VEGFR-2 rather than of both receptors may optimally overcome the adverse hematologic consequences of elevated VEGF levels found in malignancy. ..
  50. Jiang X, Cui Z, Zhu L, Guo D, Sun W, Lin L, et al. CD40-CD40L costimulation blockade induced the tolerogenic dendritic cells in mouse cardiac transplant. Int Surg. 2010;95:135-41 pubmed
    ..IL-10(high)IL-12(low) DCs with immature phenotype were induced in the tolerant recipients with the blockade of CD40-CD40L costimulation, and they obtained the tolerogenic function. ..
  51. Davis P, Nadler S, Stetsko D, Suchard S. Abatacept modulates human dendritic cell-stimulated T-cell proliferation and effector function independent of IDO induction. Clin Immunol. 2008;126:38-47 pubmed
    ..Taken together, these data aid our understanding of the mechanism for efficacy of abatacept in patients with autoimmune disease. ..
  52. Benwell R, Hruska J, Fritsche K, Lee D. Double stranded RNA- relative to other TLR ligand-activated dendritic cells induce extremely polarized human Th1 responses. Cell Immunol. 2010;264:119-26 pubmed publisher
    ..Collectively, these results demonstrate that dsRNA-activated DCs induce more highly polarized human Th1 responses than the other TLR ligand-activated DCs tested here. These results have implications for TLR ligands in immunotherapy. ..
  53. Ceballos A, Sabatte J, Nahmod K, Martinez D, Salamone G, Vermeulen M, et al. Sphingosylphosphorylcholine activates dendritic cells, stimulating the production of interleukin-12. Immunology. 2007;121:328-36 pubmed
    ..Taken together, our results support the notion that the accumulation of SPC in peripheral tissues during the course of inflammatory processes may favour the development of T helper type 1 immunity. ..