In Vitro Xenorecognition of Adult Pig Pancreatic Islet Cells by Splenocytes From Nonobese Diabetic or Non-Diabetes-Prone Mice1

As, Rivereau; You, Sylvaine; Lalain, Sandrine; Gouin, E.; P, Sai

doi:10.1097/00007890-199809150-00015

Cited by 14 publications

(19 citation statements)

References 26 publications

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“…Even though lymphocyte proliferation in diabetic and healthy subjects seemed to involve similar mechanisms with respect to blockage by MoAbs, responses of purified T cells and cytokine production, the system involving co-incubation with adult PIC could have induced more intense lymphocyte proliferation in some diabetic patients indicative of an ªautoimmuneº response. In this respect, we previously observed a higher proliferative response of splenocytes from diabetes-prone non-obese diabetic (NOD) mice than control mice to the same PIC [18]. This tendency is also consistent with the results of the only previous in vitro study in humans on this subject [19], which used fetal instead of adult pig islets.…”

Section: Discussionsupporting

confidence: 90%

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Intensity and mechanisms of in vitro xenorecognition of adult pig pancreatic islet cells by CD 4 + and CD 8 + lymphocytes from Type I diabetic or healthy subjects

et al. 1999

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Type I diabetes could be treatable by a graft of pig islets [1]. Rejection of this discordant xenograft remains, however, a major problem. Despite advances in the understanding of hyperacute rejection that have made short-term engraftment of porcine tissue a feasible objective, cell-mediated rejection can occur. This cellular rejection could be a serious problem in the case of islets, which are possibly less susceptible to hyperacute rejection [2,3].In vitro human cellular recognition in the discordant human-pig situation has been mainly investigated with stimulator pig lymphocytes or endothelial cells [4±6], i. e. cells which are not intended to be grafted in diabetes. In terms of the disease which islet grafts are intended to combat, it is thus important to characterise human anti-pig islet response since islets have particularities different to lymphocytes, e. g. they do not constitutively express class II molecules Diabetologia (1999) Summary The intensity and mechanisms of cell-mediated rejection of pig islet cells were studied in 49 Type I diabetic and 34 healthy subjects. Human peripheral mononuclear cells proliferated strongly in response to pig islet cells (p < 0.001), though with notable interindividual variations (stimulation index 2 to 215). The variance of stimulation index was higher in diabetic than healthy subjects (p < 0.0001). The response to islet cells was stronger (p < 0.01) than that to pig splenocytes. Proliferation in response to islet cells was strongly decreased (p < 0.01) when CD 4 + T cells were blocked with monoclonal antibodies, whereas the blocking of CD 8 + cells or NK cells gave less pronounced effects. The response to islet cells was decreased (p < 0.01), but not abolished, after antigen-presenting cells were removed. Purified CD 4 + cells alone did not proliferate in response to islet cells but recovered their proliferative ability when mixed with antigen-presenting cells, whereas CD 8 + cells alone proliferated in the presence of interleukin-2 in response to islet cells. Proliferation was blocked (p < 0.01) by anti-DR monoclonal antibodies. During proliferation in response to islet cells, interleukin-10 increased 43-fold (p < 0.01) but interferon-g increased only slightly. No statistical differences were detected between diabetic and control subjects with respect to lymphocyte subsets and the recognition mechanisms or to interferon-g / interleukin-10 production in response to islet cells. These results provide the first detailed information on human cell-mediated xenoreaction to pig islet cells. This situation involves a dominant CD 4 class II-restricted Th2 response, with an indirect recognition pathway, as well as a CD 8 T-cell response resulting from direct recognition. This strong reaction constitutes a serious obstacle which may vary in degree among subjects. [Diabetologia (1999) 42: 330±335]

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Section: Discussionsupporting

confidence: 90%

“…These results indicate that human cellular response to PIC differs from that of mice to the same PIC reported in our previous studies [18]. This difference has already been noted for other porcine tissues than islets [11].…”

Section: Discussionsupporting

confidence: 81%

Intensity and mechanisms of in vitro xenorecognition of adult pig pancreatic islet cells by CD 4 + and CD 8 + lymphocytes from Type I diabetic or healthy subjects

et al. 1999

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“…This is because mouse T-cells from a variety of inbred strains fail to mount primary in vitro responses to xenogeneic stimulators due to cross-species molecular incompatibilities that serve to limit the efficiency of direct xenorecognition (32). Even in those studies where a primary response to xenogeneic islets has been documented in vitro, it has often been shown to be dependent on the presence of recipient APCs, implying either a reliance on transcostimulation or an indirect presentation of processed xenoantigens (33). However, as we have previously shown (19) and confirmed again in this study, CD4…”

Section: Discussionmentioning

confidence: 99%

Achieving Permanent Survival of Islet Xenografts by Independent Manipulation of Direct and Indirect T-Cell Responses

et al. 2005

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Recent success in pancreatic islet allotransplantation has raised expectations but has equally highlighted the acute shortage of donor tissue. The use of xenogeneic tissue would help to address this shortage; however, strong cellular immunity limits the application of this approach. T-cell responses to xenogeneic tissues involve recognition of intact species-mismatched major histocompatibility complex (MHC) molecules, the direct pathway, and xenogeneic proteins presented as peptides by responder-type MHC molecules, the indirect pathway. In this study, we exploited the species difference to selectively and sequentially inhibit direct and indirect xenoresponses after transplantation of porcine islets into mice. Selective inhibition of the direct response was achieved using porcine CTLA4-Ig, which binds preferentially to pig versus mouse B7 molecules. Selective inhibition of the indirect response was achieved using murine CTLA4-Ig, which binds preferentially to mouse B7 molecules. Administration of porcine CTLA4-Ig alone caused modest prolongation of islet survival. Injection of murine CTLA4-Ig alone had a minimal effect. However, the injection of the porcine fusion protein early and the murine homolog late after grafting led to permanent survival of the porcine islets, in the absence of any other immunosuppression. These results suggest that a similar approach could have clinical utility in porcine islet xenotransplantation.

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“…The primers used for PCR amplification of PERV sequences or pig mt DNA were synthesised by the Life Tech-During xenograft, there is a risk of transmitting infectious agents from pig to man. The risk of conventional zoonosis led us to isolate islets from specific pathogen-free (SPF) pigs [1,2,3,4,5,6,7,8]. However, there is also a risk of transmitting porcine endogenous retroviruses (PERV) [9,10,11,12].…”

Section: Pcr-derived Monitoring Of Perv Infection and Microchimerism mentioning

confidence: 99%

Microchimerism and transmission of porcine endogenous retrovirus from a pig cell line or specific pathogen-free pig islets to mouse tissues and human cells during xenografts in nude mice

et al. 2002

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Aims/hypothesis. Pig islets could transmit porcine endogenous retroviruses (PERV) to diabetic patients. Our previous work showed that pig islets expressed low levels of PERV mRNA and were not likely to transmit PERV to human cells in vitro. The real risk of infection during pig tissue xenografts can only be evaluated by in vivo experiments. Methods. Nude mice bearing tumours containing human 293 cells were grafted with specific pathogenfree pig islets or PERV-producing pig PK15 cells to determine whether pig cells could transmit PERV to mouse and human cells in vivo. Infection was monitored by PCR, long PCR, RT-PCR and long RT-PCR. As detection of PERV sequences could be due to the presence of residual pig cells, we looked for pig mitochondrial (mt) DNA. Quantitative PCR for PERV and pig mt DNA was done to compare the PERV-topig mt (P-to-M) ratio of each sample with the reference ratio for grafted pig cells. Results. Among 78 mouse tissues from PK15-grafted mice, 54 and 72 were positive for gag and pig mt DNA, respectively. Human tumours developed in these mice were positive for PERV (78%) and pig mt (89%). The P-to-M ratios for mouse tissues and PERV-positive human tumours from PK15-grafted mice were higher than the ratio in PK15 cells. Among 41 tissues from pig islet cell-grafted mice, 7 were positive for PERV (3 lymph nodes, 1 kidney, 2 salivary glands, 1 ovary), and 14 were positive for pig mt DNA. Three of these samples (1 lymph node, 1 kidney and 1 salivary gland) were positive for gag DNA, but negative for pig mt DNA. One human tumour in these mice was positive for PERV DNA. P-to-M reference ratio in grafted islet cells was 0.05±0.03. The three PERVpositive lymph nodes contained 78 gag/3 mt copies (P-to-M: 26), 101 gag/3 mt copies (P-to-M: 34), and 4 gag/0 mt copies. The two PERV-positive salivary glands contained 14 gag/1 mt copies, and 28 gag/0 mt copies. The ovary and the kidney contained 46 gag/ 3 mt and 69 gag/0 mt copies, respectively. The PERV-positive human tumour contained 47 gag/3 mt copies. Conclusions/interpretation. Microchimerism and PERV transmission were frequently observed in both mouse and human tissues during grafting of pig PK15 cells into nude mice bearing human tumours, and sometimes during pig islet xenograft in this model. This strengthens the notion that there is a risk of transmitting PERV during pig islet xenograft. [Diabetologia (2002) 45:914-923] Keywords Xenograft, pig endogenous retrovirus, pig islet cell, PK15 cells, specific pathogen free pig, human cells, mouse. vided by E. Gouin (Ploufragan and Nantes, France) from Large-White SPF pigs (80-120 kg), aged 20 weeks, as previously described [1,2]. Pancreases were removed in betadine solution and inflated with 100 ml ice-cold sterile modified University of Wisconsin (mUW) solution and transported in mUW. Pancreases were then inflated again with 0.5 ml/g of mUW solution containing 2 mg/ml Liberase (Roche Diagnostics, Meylan, France) and placed in a digestion chamber filled with mUW solution kept at 36°C. Crude isle...

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In Vitro Xenorecognition of Adult Pig Pancreatic Islet Cells by Splenocytes From Nonobese Diabetic or Non-Diabetes-Prone Mice1

Cited by 14 publications

References 26 publications

Intensity and mechanisms of in vitro xenorecognition of adult pig pancreatic islet cells by CD 4 + and CD 8 + lymphocytes from Type I diabetic or healthy subjects

Intensity and mechanisms of in vitro xenorecognition of adult pig pancreatic islet cells by CD 4 + and CD 8 + lymphocytes from Type I diabetic or healthy subjects

Achieving Permanent Survival of Islet Xenografts by Independent Manipulation of Direct and Indirect T-Cell Responses

Microchimerism and transmission of porcine endogenous retrovirus from a pig cell line or specific pathogen-free pig islets to mouse tissues and human cells during xenografts in nude mice

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