Skin grafting has been one of the most important approaches for covering burn wounds, however long-term survival of allogeneic or xenogeneic skin graft is currently not successful. How to induce immune tolerance for life-time survival of allogeneic or xenogeneic skin graft is still remote objective to be solved. However, clinicians and scientists in China have worked very hard and made great contribution to this field during the past 50 years, no matter how difficult it is. They are the respected pioneers in the understanding of immunological change in "Chinese Method" skin grafting, its local immune tolerance, immunology of pre-treatment of skin graft, etc. Herein, the most outstanding and impressive progresses in immunological responses after skin grafting in the past 50 years in China have been reviewed and presented for memory, for future and for extending a salute.
Humans ; Immune Tolerance ; Skin Transplantation ; immunology ; Transplantation, Heterologous ; immunology ; Transplantation, Homologous ; immunology

OBJECTIVETo investigate the immunologic reaction difference between xenogeneic and allogeneic acellular dermal matrix (ADM) grafting.

METHODSSplit thick skin samples harvested from healthy piglets and human volunteers who underwent losing-weight operation were processed to be xeno-ADM and allo-ADM. The ADMs overlapped with ultrathin auto-skin were employed to immediately cover the wound after escharectomy in deep burn patients. The patients were correspondingly set to be Xeno (26 cases) and Allo (10 cases) groups. Another 8 cases with deep burn wounds were grafted with only split thick autoskin (TTS) after escharectomy as control group. The tissue samples from grafted area were observed by immunohistochemistry after the grafting. The typing of immune cells in peripheral blood and grafted tissue was determined.

RESULTS(1) The CD4(+), CD45RO(+) and CD4(+)/CD8(+) cell ratios in peripheral blood in Xeno group increased slightly after the skin grafting when comparing to those in control group (P > 0.05). (2) There existed lasting inflammatory and immunological reaction in the local site of grafts in Xeno group. In addition, more than 80% of the inflammatory cells could be found to be CD3(+)/CD4(+), CD45RO(+). But CD8(+), Vs8C(+) plasmocytes and CD57(+) NK cells were found less. Furthermore, eosinophil and CD68(+)/CD4(+) foreign body megalocyte reactions could also be identified, especially in Xeno-ADM before rejection (P < 0.05 - 0.001). There was only mild inflammatory and immunological reaction during early grafting stage (within 8 post-operational weeks) in Allo-group.

CONCLUSIONThe specific immunologic reaction of human host to ADM might be participated by mononuclear cells and macrophages and presented mainly as cellular immune reaction induced by CD4(+) T lymphocytes. Furthermore, the foreign body megalocyte constructed by help T cell and macrophage might play important roles in the reaction.

Animals ; Burns ; immunology ; surgery ; Dermis ; transplantation ; Graft Rejection ; Humans ; Skin Transplantation ; immunology ; methods ; Swine ; Transplantation, Heterologous ; Transplantation, Homologous

OBJECTIVETo investigate the difference of the antigenicity of xenogenic acellular dermal matrix (ADM) implants prepared by different methods.

METHODSThe split-thickness skin sheet from swine was processed by trypsin and Triton X-100 to make xeno-ADM. Twenty-five Japanese white rabbits were divided into 5 groups, i.e. xeno-ADM(1) (conjugated with glutaraldehyde), xeno-ADM(2) (conjugated with network) and xeno-ADM(3) (no conjugation, as control), in which the ADMs were and xeno-ADM(4) (conjugated) and allo-ADM (no conjugated as control), in which the ADMs were embedded into the subcutaneous place of rabbit ear and back after that the rabbits were pre-sensitized by xeno-ADM soluble protein antigen injections. The titers of anti ADMs antibody in rabbit serum were monitored during 2 - 32 post-operative weeks and the histological changes of the embedded ADMs were observed grossly and microscopically.

RESULTSThe serum titers of anti-xeno-ADM in xeno-ADM(4) group was the highest. Whereas regardless of the sensitizing effects, the titers in all groups ranged as follows: xeno-ADM(3) > xeno-ADM(2) > xeno-ADM(1) (P < 0.05 - 0.01). About 40% serum samples in allo-ADM group exhibited positive anti-allo-ADM protein antibodies. Histologically, Evident and lasting inflammatory reaction could be found in the xeno-ADM grafting sites, which was much stronger than that in allo-ADM group. The degradation and absorption gradient of ADM was ranked as follow: xeno-ADM(3) > xeno-ADM(2) > xeno-ADM(4) > xeno-ADM(1) > Allo-ADM. Foreign body megalocytic reaction might evoke in the surrounding of conjugated ADM.

CONCLUSIONThe immunogenicity in xeno-ADM was stronger than that in allo-ADM, which could induce the host to develop immune reaction restricted by IgG. Large sheets of degenerated ADM implants could lower down the antigen-antibody reaction and ameliorate the structural destroying and degeneration absorption of ADM induced by inflammatory immune reaction.

Animals ; Antigen-Antibody Reactions ; Dermis ; immunology ; transplantation ; Male ; Rabbits ; Skin Transplantation ; Swine ; Transplantation, Heterologous

OBJECTIVETo lower down the antigenicity of heterogenous swine acellular dermal tissue, and to explore the feasibility of clinical using it as a composite graft for human patients.

METHODSSplit-thickness skin was harvested from healthy swines and then processed by two methods. The swine acellular dermal matrix (sADM) was prepared by removing cells from the skin with trypsin and Triton X-100. Then the cross-linked sADM (sADM(1)) and non-cross-linked sADM (sADM(0)) were embedded subcutaneously in rabbits and also transplanted onto the burn wounds of patients. The histological changes and also transplantation results were observed.

RESULTS(1) In animals with sADM(0) embedded subcutaneously, the grafted tissue was invaded immediately by host cells with obvious inflammatory reaction and tissue degradation. But there was less inflammatory reaction, and with no obvious skin degradation and contraction with sADM(1). (2) In ten burn patients with III degree burn wounds and one patient with wound in chest after scar removal, sADM and ultra-thin skin (UTS) composite graft were grafted on the wounds with autologous thin skin (ATS) and autologous razor-thin or UTS as the control. Nineteen pieces of composite skin of sADM with UTS were grafted on the wounds with survival rate of 78.9%, exhibiting no evident difference with that of ATS. When sADM(0) and UTS were grafed, there exhibited remarkable early inflammatory reaction and wound contraction with similar external appearance with that of UTS. Whereas when sADM(1) and UTS were grafted, there appeared less early inflammatory reaction and wound contraction, resulting in an even appearance and soft to touch similar to that with ATS. But ulceration occurred, with exposure of sADM(1), exposure and severe macrophage reaction to foreign body in 6 wounds of 3 cases 12.8 +/- 6.9 weeks after sADM(1) and UTS grafting.

CONCLUSIONGrafting of sADM as a dermal substitute of composite skin could alleviate early post-grafting immune reaction and improve UTS grafting results. But the delayed graft rejection couldn't be avoided.

Animals ; Burns ; surgery ; Dermatologic Surgical Procedures ; Dermis ; immunology ; transplantation ; Humans ; Rabbits ; Skin ; immunology ; injuries ; Skin Transplantation ; methods ; Skin, Artificial ; Swine ; Time Factors ; Transplantation, Heterologous ; Wound Healing

OBJECTIVETo investigate the influence of escharectomy during shock stage on systemic and intestinal immune function and its mechanism in scalded rats.

METHODSNinety-six Wistar rats were employed in the study of which 8 were used as normal control group. The donor skin from the trunk in twenty-four rats were preserved in liquid nitrogen. The other 64 rats were subjected to 30% full-thickness scalding, and they were randomly divided into A (n = 24, no treatment after scalding), B (n = 24) and C (n = 16) groups. Physiological saline was intraperitoneally injected (50 ml/kg) on the 24 post-scalding hours to the rats in the B and C groups. The rats in B group underwent escharectomy during shock stage, and the excision wounds were covered with the cryo-preserved alloskin. The rats in C group received the same treatment as in B group but at 72 post-scalding hours. The change in the proliferative ability of splenic lymphocytes, the plasma and intestinal tissue content of interleukin 2 (IL-2), the contents of sIgA in intestinal mucus, and the content of DAO in the intestinal tissue were observed on 2, 4 and 8 post burn days (PBD) in A and B groups and also on 4 and 8 PBD in C group, respectively.

RESULTSThe splenocytic proliferative ability, IL-2 level in the plasma and intestinal tissue, and the sIgA content in intestinal mucus in the rats in A, B and C groups were lower than that in control group at all time points (P < 0.05). The proliferative ability of splenic lymphocytes in B group on 4 and 8 PBD and in C group on 8 PBD respectively was similar to that in control group. Whereas the IL-2 content in plasma and in intestinal tissue was higher in B and C groups than that in A group (P < 0.01). The sIgA content in intestinal mucus in B group was twice of that in C group respectively [(3.51 +/- 2.14) mg/g vs (1.40 +/- 0.64) mg/g, (3.03 +/- 0.95) mg/g vs (1.52 +/- 1.26) mg/g (P < 0.05 or P < 0.01)] on 4 and 8 PBD. The DAO activity in the intestinal tissue in A group was lower than that in control and B group (P < 0.05) on 4 and 8 PBD.

CONCLUSIONEscharectomy during shock stage might be beneficial to the recovery of the systemic and intestinal immune functions in rats with scalding injury.

Animals ; Burns ; immunology ; surgery ; Immunoglobulin A, Secretory ; immunology ; Interleukin-2 ; immunology ; Intestines ; immunology ; Male ; Rats ; Rats, Wistar ; Shock, Traumatic ; immunology ; surgery ; Skin Transplantation ; immunology ; T-Lymphocytes ; immunology

OBJECTIVETo observe the immune reaction in the mixed culture of host lymphocytes with allogenic and host endothelial cells.

METHODSThe host epithelial cells and lymphocytes from burn patients and allogenic epithelial cells were mix-cultured in different ratios, so as to simulate the local immune micro-environment of host skin island in intermingled skin grafting. In addition, the cells from normal human subjects were also mix-cultured as control. The lymphocyte cpm values were detected by (3)H-TdR and HLA molecules and T cell subgroup were determined by immunohistological technique.

RESULTS(1) The lymphocyte proliferation reaction could be effectively inhibited by the epithelial cells from burn patients but not from normal control. (2) The inhibition of host lymphocyte proliferation could not be mediated by the HLA-DQ molecules of epithelium from burn patients. (3) The positive expression rate of HLA-DR of epithelia from burn patients was evidently higher that that from normal control (P < 0.05), (4) The CD8 expression of lymphocyte in burn patients was significantly higher than that in normal control (P < 0.01), while the CD4 expression in burn patients was lower than that in normal control (P < 0.01). But there was no obvious difference of the CD3 expression between patients and normal subjects (P > 0.05).

CONCLUSIONThe lymphocyte proliferation reaction could be obviously inhibited by the host epithelium, which might be related to the specific immune state of the host lymphocytes and epithelium of burn patients.

Cell Communication ; immunology ; physiology ; Cell Culture Techniques ; Cell Division ; Epithelial Cells ; immunology ; physiology ; Humans ; Lymphocytes ; immunology ; physiology ; Skin Transplantation ; immunology

In the early developing stage of burn surgery, severe burn patients with large and deep burn wound often died of complications because of shortage of auto-skin. The method of intermingled transplantation composed of a large sheet of partial thickness allo-skin with punched holes for in laying small pieces of partial thickness auto-skin was first advocated by Chinese doctors (Ruijin Hospital) in 1960's. This intermingled transplantation method has saved many severe burn patients with extensive full-thickness burn wound. The mortality rate of severe burn patients has decreased and the survival rate has increased remarkably since the intermingled transplantation treatment method used in the burn units. In this paper we review the process of formation of intermingled transplantation and the mechanisms of success of this Chinese method in repairing the large wound surface area after eschar excision. We will focus our discussion on the low systemic immunological reaction, the effect of auto-skin islet, local immunological tolerance induced by in layed auto skin, the balance of Th1 and Th2 cells and the effects of some cytokines such as IL-10 in local immunological tolerance and etc. after intermingled transplantation.
Burns ; immunology ; pathology ; surgery ; Humans ; Skin Transplantation ; immunology ; methods ; Wound Healing

OBJECTIVETo observe the survival of hand allograft under the state of immunosuppression and the pathological changes of rejection in the recovery process.

METHODSThe biopsies of the skin, nerve, muscle, tendon and bone tissue of hand allografts during different stages from 1 day to 7 months after operation were observed using routine histological technique.

RESULTSNo significant changes due to rejection in skin, nerve, muscle and bone tissue were observed. But different degrees of weak rejective changes were found on the wall of blood vessels; in the muscle and nerve the reactions were markedly stronger than those found in skin tissues.

CONCLUSIONSThe rejection in deep tissues should be monitored in controlling the rejection of hand allograft.

Adult ; Biopsy ; Graft Rejection ; pathology ; Hand Transplantation ; Humans ; Immunosuppression ; Male ; Skin ; immunology ; pathology ; Transplantation, Homologous

OBJECTIVETo investigate the influence of local application of cytotoxic lymphocyte antigen 4-Ig (CTLA4-Ig) adenovirus on the burn wound with alloskin grafting upon the murine immune function.

METHODSSixty BALB/c mice were randomly divided into A (operation control), B (CTLA4-Ig transfection) and C (normal control) groups, with 20 mice in each group. Skin wounds (full-thickness loss) sized 1.5 cm x 1.5 cm were created on the backs of mice in A and B groups. Then the skin grafts of the same size obtained from C57BL mice were grafted into the skin wounds. 0.1 g of cross-linking polyacrylic resin (carbomer cream) without adenovirus was daubed onto the wounds in A group, and the same amount of carbomer cream with adenovirus in titers of 5 x 10(9)/L was daubed onto the wounds in B group, while no treatment was given in C group. 1 ml of 10% SRBC (sheep red blood cell) was injected intraperitoneally to all the mice of the three groups on the 1st post injury day (PID). Splenocytes from BALB/c, C57BL and Kunming mice were harvested for mixed lymphocyte culture on 7, 14, 21 and 28 PIDs. Agglutination assay was used in the same time to detect the SRBC antibody titers.

RESULTSThe reaction of murine splenocytes in B group to the donor (C57BL) splenocytes was suppressed in a specific way (P < 0.05) within 14 PIDs. There was no difference in the titers of anti-SRBC antibody among the 3 groups (P > 0.05).

CONCLUSIONLocal application of CTLA4-Ig recombinant adenovirus exhibited no influence on the murine humoral immunity, but might induce systemic and specific T cell tolerance in immunity system.

Adenoviridae ; genetics ; Animals ; Antigens, CD ; immunology ; CTLA-4 Antigen ; Immune Tolerance ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Skin Transplantation ; immunology ; Transplantation, Homologous ; immunology

OBJECTIVETo explore the feasibility of transplanting the skin from chimeric rats to rabbits.

METHODSChimeric rats were produced by transplanting the haematopoietic stem cells (HSCs) from rabbit marrows into fetal rats in uterus and followed by injecting the HSCs into the livers of the rats at newborn stage. After six weeks, the skin from chimeric rats was transplanted to the rabbits. In group A, the skin grafts from chimeric rat donors were transplanted to the HSCs donating rabbits, with the skin from non-chimeric rat to normal rabbits were used as control. In group B, the skin grafts from chimeric and non-chimeric rats were transplanted to the HSCs donating rabbits at the same time. Gross observation and the surviving time of heterogenic-skin graft were observed. The wound healing time was also recorded.

RESULTSIn group A, the surviving time and the wound healing time of non-chimeric grafts were (9.3 +/- 1.8) days and (20.9 +/- 2.1) days, respectively, while those in chimeric grafts were (15.1 +/- 2.6) and (18.5 +/-1.3) days, respectively. In group B, the surviving time and the wound healing time of non-chimeric grafts were similar to those of group A. Compared with those in non-chimeric grafts, the surviving time of chimeric grafts in both groups were prolonged (P < 0.01), and the wound healing time shortened (P < 0.05 or 0.01). Most of the wounds healed quickly after rejection of chimeric grafts, while the wounds with non-chimeric grafts were re-opened with exudation and some necrotic tissue.

CONCLUSIONImmunologic tolerance for skin graft can be induced by the skin from chimeric donors, which can prolong the surviving time of skin grafts and shorten the wound healing time.

Animals ; Immune Tolerance ; Male ; Myeloid Progenitor Cells ; transplantation ; Rabbits ; Rats ; Rats, Sprague-Dawley ; Skin Transplantation ; immunology ; Transplantation Chimera ; immunology ; Transplantation, Homologous