Donor-recipient gender difference affects severity of dry eye after hematopoietic stem cell transplantation.

PURPOSE: To determine whether the incidence rate and severity of dry eye after hematopoietic stem cell transplantation varies with donor vs recipient gender. METHODS: We limited this study to patients received bone marrow transplantation (BMT). In all, 172 patients received BMT at Keio University School of Medicine between January 2000 and May 2007. Of them, 136 recipients who survived at least 70 days were studied prospectively. We classified the 136 patients according to the gender of the donor and the recipient (group I: female to female; group II: male to male; group III: male to female; group IV: female to male). The incidence and severity of chronic graft-vs-host disease-associated dry eye were determined for each group. The donor gender was masked when we assessed dry eye and calculate the incidence. RESULTS: The incidence of dry eye was 47.4% for group I, 37.5% for group II, 58.6% for group III, and 42.9% for group IV. The percentage of patients with severe dry eye was 44.4, 50.0, 35.3, and 77.8% respectively. There was a significant difference between the percent severe dry eye/total dry eye incidences in groups III and IV (P=0.0375) (odds ratio, 7.6; 95% confidence interval, 1.00-101.01). CONCLUSIONS: Close attention must be paid to the development of dry eye in cases of female to male BMTs, because the ratio of severe/total dry eye is more common in cases of female to male BMTs than in other gender combination.

GVHD pathophysiology: is acute different from chronic?

Graft-versus-host disease (GVHD) is the major complication of allogeneic hematopoietic cell transplantation (HCT). GVHD occurs in acute and chronic forms. Acute GVHD usually manifests within 100 days following HSCT. It is induced by donor T cells responding to the mismatched host polymorphic histocompatibility antigens. Chronic GVHD generally manifests later (>100 days) and has some features of autoimmune diseases. It may develop either de novo or following resolution of - or as an extension of - acute GVHD. Chronic GVHD is also thought to be induced by donor T cells, but the nature of relevant antigens, the critical cellular subsets and the mechanisms of chronic GVHD remain less well understood. In this chapter we briefly discuss and contrast the pathophysiologies of acute and chronic GVHD.

[Influence of graft-versus-host disease on long-term survival of 26 patients with hematologic malignancies after transplantation].

BACKGROUND & OBJECTIVE: Graft-versus-host disease (GVHD) is a major complication of allogeneic hematopoietic stem cell transplantation (allo-HSCT), and also is an important factor affecting the outcome of transplantation. Some researches showed that either acute or chronic GVHD is often accompanied by graft-versus-leukemia (GVL) effect, and this positive effect is associated with the decrease of leukemia relapse and the prolongation of disease-free survival of recipients. This study was to assess the influence of GVHD on the outcome of allo-HSCT. METHODS: Twenty-six patients with hematologic malignancies received allo-HSCT from Mar. 1995 to Oct. 2005. The occurrence of GVHD, relapse of leukemia, and survival of recipients were analyzed retrospectively, and the correlations of GVHD to leukemia relapse and patients' survival were evaluated. RESULTS: With a median follow-up of 20 months (range 2-127 months) after transplantation, 20 (76.9%) patients developed GVHD, 1 of which had tumor relapsed; 3 of the 6 patients without GVHD had tumor relapsed. The relapse rate was significantly lower in the recipients with GVHD than in the recipients without GVHD (P<0.05). After transplantation, 16 patients survived disease-freely, and 10 died. Kaplan-Meier survival curves showed that the 3-year disease-freely survival rate was 60%. The disease-freely survival rate was significantly higher in the recipients with GVHD than in the recipients without GVHD (15/20 vs. 1/6, log-rank=7.30, P<0.05). Cox regression models showed a significantly decreased risk of death in the recipients with GVHD (risk ratio=0.2, P<0.05). Of the 20 recipients with GVHD, 17 achieved complete response (CR), 15 of whom survived disease-freely; no survived in the 3 patients who did not achieve CR. The disease-freely survival rate was significantly higher in the recipients achieved CR than in the recipients did not (P<0.05). CONCLUSIONS: GVHD is an important factor that may influence the outcome of allo-HSCT. Treatment efficacy of GVHD is significantly associated with the disease-free survival of recipients. Early recognition and treatment of acute GVHD is the key factor of successful treatment.

Vascularized lymph node transplantation induces graft-versus-host disease in chimeric hosts.

BACKGROUND: The role of lymph nodes (LNs) in adaptive immune responses has been the subject of extensive research. In previous studies, the surgical removal of lymph nodes from rat hind limbs prevented the development of lethal graft-versus-host disease (GVHD) after allogeneic hind limb transplantation to chimeric recipient rats. The purpose of this study was to establish the role of the cellular fraction versus the microenvironment of LNs in the development of GVHD in this model. METHODS: A rat model for vascularized LN transplantation was developed and graft-versus-host responses were compared after: 1) naive ACI LN cells were infused into Wistar-Furth (WF) rats as chimeric recipients (e.g. [ACI-->WF]); 2) vascularized WF lymph nodes were transplanted to syngeneic WF recipients; 3) nonvascularized ACI lymph nodes were transplanted to [ACI-->WF] chimeric recipients; 4) vascularized ACI lymph nodes were transplanted to [ACI-->WF] chimeric recipients. RESULTS: Transplantation of vascularized ACI lymph nodes to [ACI-->WF] chimeric recipient rats resulted in severe and sometimes lethal GVHD. In contrast, neither the infusion of purified ACI LN cells nor the transplantation of nonvascularized LNs led to GVHD in chimeric recipients. CONCLUSIONS: When introducing allogeneic cells into chimeric recipients, concomitant transplantation of the vascularized LN microenvironment makes a manifest difference between induction and absence of GVHD. This illustrates the important role of the LN microenvironment in adaptive immune responses.

Evidence for a graft-versus-mast-cell effect after allogeneic bone marrow transplantation.

Mast cell leukemia (MCL) is a rare form of aggressive mastocytosis with a reported median survival below 6 months. Casuistic reports suggest the effectiveness of allogeneic bone marrow transplantation (BMT) for MCL. However, these reports lack clear evidence for a graft-versus-mast-cell (GvMC) effect. We prospectively investigated the GvMC at different time points after allogeneic BMT and donor-lymphocyte infusions (DLI). Samples were gathered from a patient with MCL treated with allogeneic BMT from an unrelated HLA identical donor. Parameters for detection of a GvMC effect included flow cytometrical analysis of mast cell (MC) populations in peripheral blood and BM, BM smear and histology, chimerism analysis of flow cytometrically sorted BM CD117+/CD34- MC and testing for anti-mast cell reactivity of donor lymphocytes by interferon (IFN)-gamma ELISPOT. DLIs reduced MC from 5 to 0.5%. MC chimerism analysis demonstrated a complete recipient genotype after BMT, suggesting that the persistent mastocytosis was part of residual neoplastic disease. At 3.7 years after BMT, there is some evidence for relapse. In summary, BMT and DLIs attenuated the mastocytosis from an aggressive to an indolent form and may have improved the patients' prognosis. The in vitro data of our study indicate for the first time the existence of a GvMC effect.

Regulatory (suppressor) T cells in peripheral allograft tolerance and graft-versus-host reaction.

Among the mechanisms capable of inducing peripheral tolerance, regulatory (suppressor) T cells (Treg) probably play a key role in the control of both reactivity to self-antigens and alloimmune response. Augmentation or manipulation of Treg could improve organ allograft survival or control graft-versus-host disease, thus resulting in operational tolerance. The role of this immunomanipulation as one method of inducing tolerance has yet to be clearly defined.

CD28 knockout mice as a useful clue to examine the pathogenesis of chronic graft-versus-host reaction.

BACKGROUND: Injection of BALB/c or DBA/2 spleen cells into F1 C57BL/6 (B6) hybrids induces a graft-versus-host reaction (GVHR) of a chronic stimulatory type that results in clinical and pathologic manifestations that resemble the human systemic lupus erythematosus (SLE). The aim of the present study was to examine the role of a major T-cell costimulatory signal receptor, CD28, in the production of autoantibody and the development of an immune complex glomerulonephritis, which are common in SLE pathology. METHODS: For this purpose, CD28-deficient (CD28KO) mice were used for the source of donor lymphocytes. Chronic GVHR was induced by an injection of BALB/c or BALB. CD28KO donor cells into normal BCF1 mice. Serum titers of anti-dsDNA antibodies were assessed by enzyme-linked immunosorbent assay (ELISA) and major histocompatibility complex (MHC) class II antigen expression on B cells were tested by flow cytometry. In addition, depositions of immunoglobulin (Ig) were examined by direct immunofluorescence staining on frozen kidney sections. RESULTS: When (BALB/c x B6)F1 mice were injected with parental BALB/c lymphocytes, serum anti-dsDNA titer was significantly increased in association with nonspecific B-cell activation and IgG deposition in the glomerular basement membrane. In sharp contrast, none of these signs were observed in F1 mice, which were injected with CD28KO spleen cells. CONCLUSION: The CD28-mediated T-cell costimulatory pathway plays a pivotal role in the development of polyclonal B-cell activation, autoantibody production, and an immune complex glomerulonephritis. We propose that CD28KO mice are useful clues in examining the pathogenesis of experimental lupus nephritis.

Effects of the chromatographic fractions of the pig placental trophoblast on graft-versus-host reaction.

The trophoblast has a significant role in regulation of immune reactions at the materno-fetal interface by producing biologically active substances. In our previous studies five fractions with immunomodulatory activities were isolated by gel chromatography from trophoblast of pig placentas. To confirm the immunomodulatory effect of these trophoblast fractions on allogeneic in vivo systems and to obtain more evidence for the relevance of their activity on the maternofetal interface, their effect was studied on graft-versus-host reaction (GVHR). To assess the GVHR, the primary and secondary popliteal lymph nodes assay was used in mice. In the primary GVHR, 100 microg protein of Fraction 2-5, mixed with 5 x 10(6) allogeneic spleen cells (C57BL/6), were injected into one of the foot pads of recipient (BALB/c) mice. The secondary GVHR was induced in F1 (BALB/c x C57BL/6) mice by injection of spleen cells of BALB/c mice intraperitoneally preimmunized with allogeneic cells. The GVHR was measured by the weight of lymph nodes and by the lymphocyte proliferation. Flow cytometric analyses of the cells in the nodes with GVHR and under the influence of Fraction 4 or 5 were performed using monoclonal antibodies. In the primary GVHR, Fraction 4 or 5, injected simultaneously with allogeneic spleen cells, significantly suppressed the lymph nodes reactivity. Fractions 4 and 5 inhibited the ability of the spleen cells of mice intraperitoneally preimmunized with allogeneic cells to induce secondary GVHR in F1 mice. The Fraction 2 and 3 had no effect on GVHR. The results revealed that a group of proteins with Mr 37-7 kDa, isolated from trophoblast of pig placenta, strongly suppressed popliteal lymph node reactivity in the primary and secondary GVHR. The data provide convincing evidence for these fractions in vivo activity, for their effect across the species barrier and suggest the relevance of the same reactions on the materno-fetal interface.

Specific immunosuppression by postoperative infusion of allogeneic spleen cells: requirement of donor major histocompatibility complex expression and graft-versus-host reactivity.

BACKGROUND: Donor leukocytes may exert positive immunoregulatory effects on allograft acceptance. Most recent studies have focused on pretreatment protocols. In this study, the effect of postoperative infusion of donor leukocytes on graft survival and the phenotypic and functional requirements for infused cells were investigated in fully major histocompatibility complex (MHC)-mismatched rat heart transplant models. METHODS: LEW (RT1l) heart grafts were implanted heterotopically into abdomens of LEW.1W (RT1u), and different types of cells were infused postoperatively. Immunohistochemistry was used to evaluate histopathological changes of grafts. RESULTS: In the absence of any immunosuppressive agents, a single dose of viable donor spleen cells (SC), but not bone marrow cells, was able to prolong heart allograft survival to about 21 days, while they were rejected promptly at day 7 in controls. Infusion of T cell-depleted donor SC, irradiated donor SC or third-party (BN) SC showed no effect on graft survival. Compared with resting cells, neither in vitro nor in vivo prestimulation of infused donor SC improved graft survival. Clinical signs of graft-versus-host reaction were not observed in all above groups. Histology showed remarkable reduction in the severity of graft infiltrate and interleukin-2 receptor-positive cells in grafts of cell-treated animals. Postoperative infusion of SC of F1 generation between different strain combinations showed two requirements for infused cells to be effective: (1) expression of donor-type MHC antigens and (2) strong alloreactivity against the host MHC antigens. CONCLUSION: Postoperative infusion of viable donor SC can lead to allospecific down-regulation of alloreactivity by a graft-versus-host-associated effect.

Regulatory function of factor-XIIIa-positive dendrocytes in incipient toxic epidermal necrolysis and graft-versus-host reaction. A hypothesis.

BACKGROUND: Lymphocyte-poor graft-versus-host-reaction (GVHR) and toxic epidermal necrolysis (TEN) share some histological resemblance. In both diseases, factor-XIIIa-positive dendrocytes show some morphological changes, probably as a response to altered cytokine environment. OBJECTIVE: To study the ultrastructural aspect of boosted dendrocytes in GVHR and TEN. METHODS: Sixty GVHR and 25 TEN lesions were examined using immunohistochemistry. Among them, 6 dendrocyte-rich cases of each disease were studied by electron microscopy. RESULTS: Dendrocyte activation with enlarged endoplasmic reticulum, and collagen fiber and mast cell granule phagocytosis were evidenced in both diseases. Depletion in dendrocytes was only encountered in a few GVHR cases exhibiting specifically a sclerotic aspect in the superficial dermis. CONCLUSION: Factor-XIIIa-positive dendrocytes probably play a role in the regulation of the connective tissue remodeling that may accompany epidermal destruction.

Low levels of eicosapentaenoic and docosahexaenoic acids mimic the effects of fish oil upon rat lymphocytes.

Fish oil is rich in the long chain n-3 polyunsaturated fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA); typically these fatty acids constitute 20 to 25 g/100 g total fatty acids in fish oil. Feeding rodents diets rich in fish oil has been shown to decrease lymphocyte proliferation and natural killer cell activity. It is not known what level of EPA + DHA is required in the diet to exert these effects. This question was addressed in the current study. Weanling rats were fed on high fat (178 g/kg) diets which contained 4.4 g alpha-linolenic acid (control) or 4.4 g EPA + DHA (4.4 EPA + DHA) or 6.6 g EPA + DHA (6.6 EPA + DHA)/100 g total fatty acids. The n-6 to n-3 polyunsaturated fatty acid ratio was maintained at approximately 7. The fatty acid compositions of the serum and of spleen leukocytes were markedly influenced by that of the diet. Spleen lymphocyte proliferation in response to concanavalin A, spleen natural killer cell activity and PGE2 production by spleen leukocytes were reduced by feeding the EPA + DHA diets compared with feeding the control diet; the 4.4 and 6.6 EPA + DHA diets caused very similar reductions. The 4.4 EPA + DHA diet reduced popliteal lymph node weight following a localised graft versus host response; this response was not investigated in rats fed the 6.6 EPA + DHA diet. The reductions in lymphocyte functions and in the in vivo graft versus host response caused by the EPA + DHA diets were similar to those previously reported following the feeding of diets rich in fish oil. Thus, this study shows that diets containing relatively low levels of EPA + DHA (20 to 25% of the level found in fish oil) exert immunomodulatory effects. Furthermore, this study suggests that the maximal effect of EPA + DHA is exerted when these fatty acids constitute a level of less than or equal to 4.4 g/100 g total dietary fatty acids.

The resorption of bone-implanted corals varies with porosity but also with the host reaction.

Three different exoskeletons of coral species Porites asteroides (P), Montastrea annularis (M), and Dichocoenia stokesi (D) were implanted for 2-20 weeks in rabbits. At 2, 4, 8, or 20 weeks, the exoskeletons presented variations in their resorptions depending on the species. To understand the variations in the decreasing speed of the implants despite their similar chemical composition, a study of the surface and architecture of the coral was carried out using scanning electronic microscopy, porosity was evaluated, and growth and differentiation of osteogenic cells cultured in vitro were observed for more than 1 month. At the cellular level, the surface of the implants was identical. Three-dimensional structures of the implants were variable, but the porosity values [P = 42.7%, M = 40.7%, and D = 17.4%] could not completely account for the differences in the resorbing process of the species. Standard histologic studies performed at 2, 4, 8, and 20 weeks after implantation produced the same pattern with P or M, showing aspects of rapid resorption; however, with D there were images resembling those of a foreign-body reaction. It seems that when resorption is not quick enough, a foreign body reaction develops which further slows down the process. This work focuses on the importance of porosity when using coral as bone substitute.

Age-related changes in graft-versus-host reactivity of spleen cells from male, virgin, or multiparous female Long-Evans rats.

Spleen cell graft-versus-host (GVH) reactivity was determined in male and female, either virgin or breeder, Long-Evans (LE) rats from 3 to 24 months of age. The tests of a regional (popliteal lymph node enlargement index) and a systemic (splenomegaly index, mortality assay) GVH reaction was used. Although the GVH reactivity declined with age in both sexes, the onset of this decline was significantly delayed in 18-month-old virgin females in comparison with 12-month-old males. Moreover, 6 or 7 consecutive pregnancies resulted in significantly enhanced GVH reactivity of 18-24-month-old females. This long-lasting effect of multiparity was observed in females mated either syngeneically or allogeneically. The possible role of neuroendocrine factors in delaying the age-related process of thymic involution in multiparous females is suggested.

Reappraisal of histologic features of the acute cutaneous graft-versus-host reaction based on an allogeneic rodent model.

We employed a rat model of complete major histocompatibility complex-mismatched allogeneic bone marrow transplantation to better characterize the histologic expression of the acute cutaneous graft-versus-host reaction (GVHR), compared with changes due to the preparative regimen. Cyclosporin A abolished the development of this GVHR. Low levels of dyskeratotic cells were present in all groups (allogeneic and syngeneic transplants with and without cyclosporin A) and, alone, were insufficient to diagnose a cutaneous GVHR. A consistent histologic feature of the GVHR was significant lymphoid infiltration of the dermis. The pattern of cytotoxic folliculitis involved follicular epithelium above the entry of sebaceous glands. Immunostain for major histocompatibility complex class II, IA, and IE antigens revealed that dendritic cells within the follicle were limited to this upper region and that lower follicular epithelium did not upregulate expression with evolution of the GVHR. Based on this model, we conclude 1) that the diagnostic scheme for the acute cutaneous GVHR should include lymphoid infiltration of the dermis, 2) that the preparative regimen (including total body irradiation) induces persistent low levels of dyskeratotic cells (two to three cells/linear mm of epidermis), and 3) that the pattern of follicular involvement may relate to the distribution of dendritic cells and to an inability of lower follicular epithelium to upregulate major histocompatibility complex class II antigens.

Biology of acute and chronic graft-versus-host reactions: predictive value of studies in experimental animals.

Experimental research on graft-versus-host disease (GVHD) with laboratory animals has been performed mainly with rodents, rhesus monkeys and dogs. The basic immunological mechanisms operative in GVHD are largely similar in these three species and in human patients, although the patterns of GVHD in the three animal species show differences. The predictive value for clinical GVHD of the results obtained in the different animals species is analysed for the three main variables: namely, histocompatibility, T cell numbers in the graft and the intestinal microflora. Rhesus monkeys score highest as regards clinical relevance for the first two variables. With regards to the unravelling of detailed mechanisms of the influence of the microflora, none of the three animal species is likely to provide the information needed for identification of the bacterial species involved in the induction of GVHD in human patients.

Embryonic Purkinje cells grafted on the surface of the adult uninjured rat cerebellum migrate in the host parenchyma and induce sprouting of intact climbing fibres.

By grafting solid pieces of cerebellar anlage onto the surface of the adult rat cerebellum, we have investigated the problem of the interactions between embryonic and adult neurons in an intact brain. A few days after grafting, embryonic astrocytic processes crossed the graft--host interface and radiated into the recipient molecular layer. Several grafted Purkinje cells also migrated into the host brain along such processes as well as adult Bergmann glia. Adult climbing fibres, labelled by means of Phaseolus vulgaris leucoagglutinin (PHA-L), sprouted new collateral branches which terminated on embryonic Purkinje cells at both extra- and intraparenchymal levels. No sign of activation of host astroglia or microglia was evident in the host cerebellum in relation to these processes. Embryonic Purkinje cells which migrated into the host cerebellum developed an adult-like morphology. Intraparenchymal grafts of neocortical embryonic tissue induced conspicuous growth of host olivary axons, characterized by a pattern which was different from that observed following cerebellar grafts. By contrast, when neocortical tissue was placed onto the surface of the recipient cerebellum, graft--host interactions were limited and climbing fibre sprouting was rarely seen. These results show that (i) supernumerary Purkinje cells can penetrate and settle in the adult intact cerebellar cortex, (ii) adult climbing fibres are able to innervate these new targets in the absence of any injury or activation of non-neuronal cells of the adult brain, and (iii) in the absence of damage to the adult brain, the plasticity of adult olivary axons is specifically elicited and controlled by embryonic Purkinje cells.