Inhibitory effect of galanin on dopamine induced increased oxytocin secretion in rat neurohypophyseal tissue cultures.

The regulation of oxytocin (OT) release by galanin (GAL) at the neurohypophyseal (NH) nerve terminal is not adequately understood. The effect of GAL on the secretion of OT was studied in 13- to 14-day cultures of isolated rat NH tissue. By this time, the hormone content of the medium had become constant. The OT content of the supernatant medium was determined by RIA after a 1- or 2-h incubation. A significantly decreased content of OT was found following incubation with 10(-6)-10(-8) M doses of GAL. Dopamine (DA) and the DA-active drugs apomorphine (APM) and Pro-Lys-Gly (PLG) (10(-6) M in each medium) increased the OT synthesis of NH tissue cultures. This elevation of OT secretion could be blocked by the administration of GAL together with DA, APM or PLG. The DA-blocking effect of GAL was prevented by previous treatment with the GAL receptor antagonist galantid (M15). The results indicate that OT release from the NH is directly influenced by the GAL-ergic system. The GAL-ergic control of OT secretion from NH tissue in rats can occur at the level of the posterior pituitary.

Importance of plasma cells in the infiltrate of renal allografts. An immunohistochemical study.

The cellular infiltration in 42 needle and wedge biopsies of transplanted kidneys was investigated immunohistochemically. The percentages of helper/inducer (CD 4+) cells, suppressor/cytotoxic cells (CD 8+), B lymphocytes, macrophages, plasma cells (Pc) and granulocytes were determined. The proportions of the various inflammatory cell populations were established in acute interstitial rejection (AIR), acute vascular rejection (AVR), chronic rejection (CR) and cyclosporin A nephrotoxicity (CsAN). The most prominent differences were detected as regards the Pc, whose number was much higher in CR than in AIR, AVR or CsAN. The striking difference between CR and CsAN in the number of Pc may be of differential diagnostic importance: the presence of many Pc in the biopsies can be regarded as a sign of CR. Over 80% of the Pc in CR contained IgG, whereas in chronic interstitial nephritis (CIN) the IgA-positive Pc predominated. In AIR, AVR and CsAN, too, the majority of the Pc contained IgG, but the numbers of IgM and IgA-positive cells were also relatively high. The great number of IgG-positive Pc indicates an important role of a secondary type humoral immune response in CR.

[Comparative analysis of ultrasonic studies, histopathological data and clinical parameters in dysfunctioning renal transplantation]. / Ultrahangvizsgálatok, kórszövettani eredmények és klinikai paraméterek összehasonlító elemzése transzplantált vese diszfunkció esetén.

The correlation of B mode and Doppler sonographic parameters and diagnoses established by histological examination of graft biopsies, nephrectomies and clinical data are discussed. 48 histological samples from 36 patients were reevaluated. The maximum interval between sonography and histology was 36 hours. The Banff classification criteria were used during histological examinations. Doppler examination evaluation was based on the resistance index (RI). Reproducibility was controlled by means of intra- and interobserver variability in 10 patients. RI values higher than 75% were regarded as abnormal. On the basis of these observations and the literature data specific sonographic features can be detected in renal artery occlusion and renal vein thrombosis. In pyelonephritis, dilatation of the collecting system was frequent. No morphological changes were detected in cyclosporin-A nephrotoxicity and the Doppler signs were not characteristic for this disease. No differentiation was found between acute rejection and acute tubular necrosis. The noninvasive duplex sonographic examinations can provide very important information regarding the flow situation of a transplanted kidney. In some cases a definitive diagnosis can be achieved, but in other cases biopsy is the method of choice.

[The role of ultrasonography and biopsy following kidney transplantation]. / Ultrahangvizsgálat és biopszia szerepe és helye vesetranszplantáció után.

Several complications can occur during both the early and late postoperative periods after kidney transplantation. The methods used to follow up 575 kidney transplanted patients, (transplantations performed between October 1979 and November 1997) in the early (within 6 weeks) and late postoperative periods have been assessed. The diagnostic value of core biopsies and ultrasound examinations, the prevalence of complications, and the applicability of the diagnostic tools in the evaluation of the graft status and viability were analyzed. In the early postoperative period, graft rupture occurred more frequently after biopsy than in the late period (7.4% vs 0.82%), this leading graft loss in 18 of 20 cases. The sonographically diagnosed morphologic and functional changes were also analyzed. Sonography proved a very accurate method for the detection of perirenal fluid collections and masses and severe vascular complications. The data demonstrated that biopsy is indicated in the early postoperative period when the result of sonography is doubtful. In the late postoperative period, biopsy should be performed in every case.

[Recurrence of focal sclerosing glomerulonephritis in the kidney graft]. / Focalisan sclerotisáló glomerulonephritis recidívája vesetranszplantátumban.

This is the first report on the recurrence of a glomerular disease in renal transplant in Hungary. The primary disease of the girl died at the age of 13 was focal sclerosing glomerulonephritis with slight mesangial cell proliferation. The first symptoms appeared at the age of 6.5 and they progressed rapidly. Four years later, because of the severe nephrotic syndrome and chronic renal failure, renal transplantation was performed with the synchronous removal of the patient's own kidneys. In the latter an interesting immunohistological finding has been observed: beside the usual positivities, the basement membrane of the distal tubule at the opposite side of the macula densa showed a strong reaction with anti IgM and a somewhat weaker positivity with anti C3 sera. The primary disease recurred very soon. A mesangial cell proliferation, however did not develop, in contrary to the primary disease, which contradicts the theory that the mesangioproliferative form would be a distinct clinicopathological entity.

[Characteristics of cellular infiltration in rejected renal grafts]. / A sejtes infiltráció jellemzése a kilöködö vesetransplantatumban.

The composition of the cellular infiltrate in 42 needle and wedge biopsies of transplanted kidneys was investigated immunohistochemically. The various inflammatory cell populations were examined in different rejection types and cyclosporin-A nephrotoxicity (CsAN) as well as in different locations in the graft (perivascular and intertubular area, tubular epithelium, glomeruli) separately. There was generally a Th cell predominance except the most unfavorable rejection type, the acute vascular rejection (AVR), where the Tc cells outnumbered all other infiltrating cell populations. The most macrophages too were detected in AVR. The high proportion of plasma cells in chronic rejection indicate an important role of the humoral immune response in this type of rejection, and could also be used as a differential diagnostic sign versus CsAN.

In-vitro steroid sensitivity in patients with chronic uraemia and renal transplantation: possible association with HLA phenotypes.

We studied in-vitro steroid sensitivity using the test of ADCC in 207 haemodialysed chronic uremic patients, 85 renal transplant patients, and 75 healthy blood donors as normal controls. The association of HLA phenotypes with the in-vitro steroid sensitivity was assessed. The proportion of steroid-sensitive subjects was significantly higher in the normal control group than in the patients. A significant association was observed between HLA B8 carriers and steroid resistance and between HLA DR6 carriers and steroid sensitivity.

Hematuria of tubular origin.

Hematuria (HU) of tubular origin is described on the basis of ultrastructural investigations of two rejected human renal allografts. The red blood cells (RBC) got into the interstitium through the walls of the damaged intertubular capillaries, and, via ruptures of the tubular basement membrane (BM), they may pass from the interstitium into the tubular lumen, causing HU. Rupture of the tubular BM may develop owing to the enzymes released by interstitial inflammatory cells or to the difference in the interstitial and intratubular pressures. Some of the RBC already in the tubular lumen may penetrate between the epithelial cells and even between the epithelium and the BM, probably because of the locally increased intraluminal pressure. If the RBC reach the BM, the epithelial cells produce a new, thin BM above them, with disintegration of the outer older one, and, thus, the RBC may pass into the interstitium again. This phenomenon is called the tubulointerstitialis circulation of RBC.

DR beta-RFLP analysis of serologically DRw6 compatible kidney donor/recipient pairs.

HLA-DR beta-RFLP analysis of serologically 1 or 2 DR-antigen compatible renal allograft donors and recipients (D/R), selected for DRw6 antigen (study group), or for other DR antigens (control group) and its correlation with the graft outcome revealed that: 1. In the study group where donors and recipients were selected for matching in DRw6 antigen, 8 out of ten D/R pairs turned out to be incorrectly classified due to a number of mismatches in the HLA-DR locus RFLP analysis. In the control group, only 2 out of ten patients matched poorly by means of RFLP. The difference between the two groups was significant. 2. A significantly higher incidence of rejection episodes was seen in the study group (2.7 +/- 1.5) than in the controls (1.4 +/- 0.5). 3. The total dose of methylprednisolone necessary for suppressing the rejection episodes was higher in the study group (6.0 +/- 2.9 g) than in the control one (4.07 +/- 2.77 g), but the difference was not significant. 4. The graft survival 2 years after transplantation was 30% in the study group and 70% in the control group.

Tubular ultrastructure in rejected human renal allografts.

Twenty percutaneous renal transplant biopsies and 20 removed allografts were investigated ultrastructurally. Most of the detected alterations were of a degenerative or regenerative nature and not specific of rejection. The most interesting phenomenon was the tubulitis, namely, the migration of the interstitial inflammatory cells (IC) through the tubular basement membrane (BM) and the invasion of the tubular epithelium in this way. Tubular epithelial cells (TEC) in the vicinity of IC were often necrotic. The composition of cells invading the tubules corresponded to those infiltrating the interstitium. The distal tubule was more frequently infiltrated than the proximal tubule. The TEC were always in very close contact with the BM. The invading IC were in direct contact with the inner surface of the BM only while passing through it. IC that passed the BM were immediately separated from it by a thin epithelial layer. The tubular ultrastructural changes did not reveal substantial differences between the various rejection types, except for the pronounced thickening and lamellation of the BM in chronic rejection.

Successful simultaneous transplantation of kidney and fetal pancreatic islet masses.

This paper reports our experience with the successful simultaneous transplantation of kidney and fetal pancreatic islets in 46-year-old diabetic man. No detectable C-peptide level was noted and the end-stage nephropathy required hemodialysis. The cadaver kidney and two masses of 8-week-cultured fetal islets were grafted simultaneously. After revascularization of the kidney, the islet masses were placed under the kidney capsule. Following transplantation, islet function was demonstrated by a higher C-peptide level, which subsequently persisted. Twenty-four months after grafting, islet function was provoked by glucagon and glucose, which led to elevations in the C-peptide and insulin levels. The insulin requirement fell from 58 to 24 U/day during the post-transplant period of 24 months. The mean value of HbA1C (5.6% +/- 0.3%) indicated a constantly normal carbohydrate metabolism. Improvements in retinopathy were also noted. Three periods of kidney rejection were diagnosed, but these proved reversible with high-dose steroid treatment. The serum and urine beta-2-microglobulin levels correlated well with rejection and recovery. More than 2 years after grafting, kidney functions is in the normal range. On sonography, the transplanted islet masses were repeatedly clearly visible, and 24 months following transplantation the volume was twice the original one. The results indicate that simultaneous kidney and fetal pancreatic islet grafting is advantageous in end-stage nephropathy secondary to type I diabetes mellitus.

[Significance of plasma cells in the rejection of kidney grafts]. / A plasmasejtek jelentösége vesetranszplantátumok kilöködésében.

Authors studied with immunohistochemical methods the immunoglobulin content of plasma cells during the rejection of renal grafts, in different rejections, in Cyclosporin-A nephro-toxicity and in chronic interstitial nephritis, as a comparison. By far the most plasma cells occurred in chronic rejection, containing more than 80% IgG. Whereas great majority of plasma cells were IgG positive in chronic interstitial nephritis. Probably, in chronic rejection a special, secondary type humoral immune reaction has a significant role in addition to cellular immune mechanisms.

About DR beta-restriction fragment length polymorphism (RFLP) analysis in kidney transplantation in connection with a paediatric patient.

A retrospective analysis of the HLA-DR antigens was performed at the DR beta DNA locus by the means of restriction fragment length polymorphism (RFLP) in the case of a two-times kidney transplanted paediatric patient and in 16 adult kidney donor recipient pairs in order to prove the importance of DNA molecular analysis in those cases where the serological identification is poor. The child and her grafts (first from her mother, the second from a cadaver donor) carried the DRw6 antigen which serologically can very poorly be defined. According to DR serotyping before transplantation both the child and the cadaver kidney proved to be DR5, 6, while the DNA analysis revealed mismatches; the child possessed the two subtypes: 13b1 and 14a of the DRw6 antigen only and none of the DR5, the cadaver kidney proved to belong to the DR4 antigen group instead of DR5, and furthermore to a different subtype of the DRw6 (13a3) than the recipient. The DNA analysis of other 16 adult donor-recipient pairs also underlined the importance of the DR beta RFLP analysis in cases where the transplantation antigens could be poorly defined.

Monitoring of suppressor cell activity in nine renal transplant patients.

Function of Concanavalin A generated suppressor cells was serially tested on lymphocytes stimulated by mitogens, by a pool of test cells different in HLA-ABC and -DR, and by the kidney donor's cells preserved in liquid nitrogen. In six patients whose lymphocytes had strong suppressor cell activity, the kidney had a good function for 17-28 months (up to the time of the survey). In three patients who had practically no suppressor cell activity before and after transplantation, each kidney was rejected within 2-3 months. These observations are in accordance with recent experimental and clinical studies of other authors supporting the importance of suppressor cell activity in transplantation tolerance.

[Pathomechanism of the development of chronic obliterative transplantation arteriopathy in human kidney allografts]. / Az idült obliteratív transzplantációs arteriopathia kialakulásának patomechanizmusa humán vese-allotranszplantátumokban.

Authors examined cells participating in intimaproliferation in transplantation arteriopathy ultrastructurally in needle and wedge biopsy material from 40 transplanted kidneys, and immunohistochemically in 10 cases. In early biopsies--even in two control kidneys--it could be observed that the smooth muscle cells of media are in direct contact with endothel cells by their small processes. Processes can fulfil a receptor function and can transmit endothel noxa to smooth muscle cells. Smooth muscle cells of media react to endothel damage caused by rejection with migration to intima and during this period they are transformed to myofibroblasts (myointimal cells). In the mean time inflammatory cells (mainly macrophages, helper and cytotoxic cells in lower number) from the lumen infiltrate the intima, and mediators, enzymes released from them can inspire smooth muscle cells to further proliferation, migration to intima and transformation to myofibroblast. To effect of mediators (gamma interferon) released from inflammatory cells, the myointimal cells during rejection will press out 2nd class transplantation antigens (HLA-DR), and as vicious circle it further aggravates immune reply to graft, causing vascular damage, intimaproliferation.

Comparative distribution of laminin and collagen types I, III and IV in transplant vasculopathy.

Composition of the extracellular matrix (ECM) was studied in transplant vasculopathy occurring in rejected renal allografts using the immunoperoxidase technique with antisera against laminin, and collagen types I, III and IV. In acute transplant vasculopathy the loose ECM network of the intima showed intense immunostaining for laminin and type IV collagen. Type III collagen was detected in the advanced acute cellular intimal proliferations, while early acute lesions did not show immunreactions. Type I collagen was not seen in significant amount. In contrast to these findings in chronic transplant vasculopathy associated with intimal fibrosis the ECM was largely composed of interstitial collagen types III and I, while staining for the basement membrane type ECM components were markedly reduced. Degradation of the matrix components with variable composition was noted in foci of mononuclear infiltrates occurring inside the fibrotic intima. These results indicate that the ECM shows a compositional change in transplant vasculopathy which is associated with the age of the lesion.

Pathogenesis of transplantation arteriopathy. Myointimal cells express HLA-DR antigens during rejection.

Arteries were investigated ultrastructurally in material from 40 needle and wedge biopsies of renal allografts, and immunohistochemically in another 10 cases with signs of chronic obliterative transplantation arteriopathy. In the early biopsies, but even in the control kidneys, thin extensions of the smooth muscle cells of the media were observed, which were in direct contact with the endothelial cells through the lamina elastica interna. These extensions may contain receptors mediating endothelial noxae to the smooth muscle cells thus initiating their proliferation, migration to the intima presumably begins in the early post-transplant period and continues until the lumen is occluded. Concomitantly, inflammatory cells (mainly macrophages, with a smaller number of CD4 and CD8-positive T lymphocytes) invade the intima. The proliferating myointimal cells, possibly having become HLA-DR-positive, may behave as antigen-presenting cells, enhancing the anti-graft immune response further, and aggravating the arterial injury.

Steroid sensitivity of chronic uraemic and renal transplant patients measured by the antibody dependent cellular cytotoxicity reaction.

The steroid (methylprednisolone) sensitivity of chronic uraemic and renal transplant patients was examined on the basis of the extent of inhibition of the antibody-dependent cellular cytotoxicity (ADCC) reaction, and via the effect on the ADCC capacity test (ADCC-C). Individuals with an inhibition of 30% or more were classified as steroid-sensitive. Immunopharmacological tests and the clinical picture showed 67%, 12 of the 18 renal transplant patients to be steroid-sensitive. In 92% of the cases the transplanted kidney was functioning well one year or more postoperatively. In 5 of the 6 steroid-resistant patients rejection necessitated removal of the transplanted kidney. The method is simple to perform and gives reproducible results, and appears suitable for application in clinical practice.