Kentaro Takebe

Quantification of serum-soluble HLA class I antigens in patients with gastric cancer

The amount of sHLA-I in serum was examined in 74 patients with gastric cancer and 15 normal healthy controls. For mAbs, W6/32 specific for HLA-A, -B, -C, and biotin IOT2 specific for HLA class I associated with beta 2 microglobulin, were used to determine the values of sHLA-I using an ELISA. The patients in stage-IV gastric cancer showed lower values of sHLA-I (445.4 +/- 247.1 ng/ml) than those in stage I (725.9 +/- 575.8 ng/ml), stage II (752.8 +/- 255.0 ng/ml), and normal controls (868.9 +/- 715.0 ng/ml) (P < 0.05). In analysis of the patients with HLA-A24, the allele that has been reported to secrete more sHLA-I than other alleles, the results were nearly the same. These results suggest that the secretion of sHLA-I is low in patients with very advanced cancer. However, there was no correlation between the sHLA-I level and the metastasis or prognosis in longitudinal studies in 11 patients.

Xenogeneic (pig to rat) fetal liver fragment transplantation using macrocapsules for immunoisolation

Acute liver failure caused by viral infection, surgical resection of a large part of the liver or by drug use has a high mortality. For its treatment, hepatocyte or liver tissue transplantation is useful. We report here the beneficial effects of xenogeneic fetal liver fragment (FLF) transplantation with an immunoisolation macrocapsule. The macrocapsules were made of a microporous polypropylene membrane. Pig FLFs (1 mL) was inserted into each capsule to serve as a graft in LEW rats. Acute liver failure was induced by 90% liver resection on day 0. Group 1: transplantation of encapsulated FLF into the omentum 2 days before liver resection (n = 17). Group 2: FLF transplantation into the omentum on day -2 (n = 11). Group 3: liver resection (control) (n = 19). The survival rate, the histology of the grafts and the biochemical parameters [blood sugar (BS), GPT, and GOT] were evaluated. The survival rates of groups 1, 2, and 3 on day 7 were 70.6, 0, and 11.1%, respectively. There were significant differences in BS, GPT, and GOT levels between groups 1 and 3 on day 1 (p < 0.05). On day 28, the histological analyses of the grafts of encapsulated FLFs revealed that the hepatocytes appeared viable, but that the haematopoietic cells had degenerated. Xenogeneic FLFs with macrocapsules survived more than 1 mo, and supported the hosts liver function.

Soluble HLA class I and class II in Japanese

Soluble human leukocyte antigens (sHLA) class I (sHLAI) and class II (sHLA-II) in sera or plasma can be quantitatively measured by enzyme-linked immunosorbent assay (ELISA) [1, 2]. Several reports state that individuals with particular HLA allotypes have a higher sHLA-I concentration than those without that particular allotype [1-5]. HLA-A9 especially is widely known to be accompanied by the highest sHLA-I concentration [2-5] . Meanwhile, there are only a few reports discussing sHLA-II quantification [6, 7] and no definite conclusion about the association with particular HLA allotypes has been reported. In the present study, sHLA-I and II (DR) in plasma were measured by ELISA and their association with particular HLA allotypes was examined in Japanese subjects. The frequency of HLA-A9-positive individuals in Japan is much higher (68%) than that in North America (22%) [8]; therefore, the Japanese population could provide the appropriate samples to investigate the relationship between sHLA-I and A9 allotype.

Serum concentrations of soluble HLA-class I and CD8 forms in patients with viral hepatic disorders

Soluble HLA-class I and CD8 molecules were determined by sandwich ELISA in patients with viral-induced hepatic disorders. As a whole, the patients with hepatic disorders (acute hepatitis: AH; chronic hepatitis: CH; liver cirrhosis: LC; hepatocellular carcinoma: HCC) showed higher sHLA-class I and sCD8 levels than normal controls (P<0.001). AH patients had the highest sHLA-class I levels (mean, 3513±2112ng/ml), followed by CH (2896±1290ng/ml), LC (2293±1266ng/ml), and HCC (2221±1212ng/ml) sCD8 levels were highest in AH, followed by HCC, LC, and CH, in that order. Among histologically defined C virus-positive patients, sHLA-I levels were higher in those with chronic active hepatitis (CAH) 2A (3802±1124ng/ml) than in those with chronic persistent hepatitis (CPH; 2200±711ng/ml;P<0.01), the levels then decreased as the disease progressed (CAH2B, 3564±1783ng/ml, LC, 2376±1265ng/ml). In contrast, sCD8 values showed little difference among the disorders. sHLA-class I levels showed a positive correlation with sCD8 values both in whole patients and in patients with AH (P<0.01), but no correlation was shown, in any patients, with biochemical parameters such as GPT and GOT. These findings, taken together, suggest that hepatic destruction is not the only cause of sHLA-class I production, but that sHLA-class I levels, together with sCD8 levels, may reflect immunological activity in hepatic disorders.

Effects of iso and xeno fetal liver fragments transplantation on acute and chronic liver failure in rats.

Isogeneic (rat) and xenogeneic (swine) fetal liver fragments (FLF) transplantation into the omentum was performed for d-galactosamine (d-Gal)-induced acute and carbon tetrachloride (CCI4)-induced chronic hepatic failure in rats. The recipients that had iso or xeno FLF showed higher survival rates than the nontransplanted controls on a lethal dose (2.6 g/kg body weight) of d-Gal (survival rates: Iso 70%, Xeno 80%, and control 9.1%). On a sublethal dose (1.0 or 1.2 g/kg) of d-Gal, iso, or xeno FLF caused marked improvement of the values of GPT, GOT, and total bilirubin (T. Bil); at 72 h after d-Gal injection they went significantly lower than those of controls (Iso vs. control; p < 0.01, Xeno vs. control; p < 0.05). Histological examination of the livers revealed severe damage in controls, however, only a slight damage was found in iso or xeno FLF transplanted rats. Iso grafts were fairly well preserved in the omentum at 72 h posttransplants, however, xeno graft had almost changed into a necrotic tissue. CCI4 was administered subcutaneously for 14 wk to induce chronic hepatic failure and then iso FLF were transplanted 3 days after the last CCI4 injection. Iso FLF transplanted rats showed higher improvement of GPT and GOT values at 12 days posttransplants compared with controls (GPT p < 0.01, GOT p < 0.05), although histological improvement was not so remarkable in both group. Iso grafts formed nodules with many hepatocytes in the omentum 12 days posttransplant. The results indicate that iso or xeno FLF transplantation could be an alternative approach for incurable liver insufficiencies.

The changes of serum soluble HLA-class I and CD8 concentrations during interferon treatment of chronic hepatitis C patients

Serum soluble HLA-class I and CD8 molecules were sequentially (pre, during, and post-therapy) measured by sandwich ELISA, in 20 interferon (IFN)-treated chronic hepatitis C (CHC) patients, who were judged as responders (n = 8) or non-responders (n = 12) with respect to the response to the therapy. Pretreatment levels of sHLA-I and sCD8 did not differ between responders and non-responders. Both values increased just after the start of therapy and peaked at 2–4 weeks. The IFN was discontinued at 24 weeks, when sHLA-I decreased to baseline levels in responders, whereas they remained at levels significantly higher than baseline levels even at 48 weeks in non-responders (P < 0.01). sCD8 decreased significantly under than baseline levels at 48 weeks in responders (P < 0.05), in contrast in non-responders, sCD8 did not decrease beyond baseline levels. In summary, serum levels of sHLA-I and sCD8 were elevated by the IFN treatment, and serial follow-up of these immunological parameters could provide additive information regarding the response to IFN therapy.

Analysis of survival of allogeneic fetal liver fragments in rats

The survival of allogeneic fetal liver fragments in the omentum was analyzed in rats. The lymphocyte subsets of the spleen and peripheral blood were also examined. When the fetal liver fragments were transplanted into the omentum, they survived for 2 wk, whereas adult liver fragments survived only 1 wk. In fetal liver fragments transplantation, the CD8 positive lymphocyte percentage in peripheral blood decreased significantly 3 wk after transplantation in comparison with that in adult liver fragment transplantation. The skin graft of the donor party showed a longer median survival time in rats receiving fetal liver fragment transplants than that in recipients of adult liver fragments. Although further study is needed, allogeneic fetal liver fragments survived longer in the omentum than reported elsewhere, and the decrease of CD8 positive peripheral blood lymphocytes may have been the reason for this.

A Case Report of Anal Carcinoma Associated with Perianal Pagetoid Lesion

肛門周囲Paget病を伴う肛門癌は, きわめてまれな疾患であり, 本邦では今回われわれが経験した症例を含めて9例が報告されているにすぎない.症例は74歳男性であり, 肛門周囲の疼痛と腫瘤を主訴として入院, 皮膚生検の結果肛門周囲Paget病と診断された.さらに肛門癌も合併していた.手術は腹会陰式直腸切断術を行ったがすでに両凧径部リンパ節転移を認めた.術後, 会陰部創に対しては計20Gyの腔内照射, 嵐径部に対しては計14Gyの電子線照射を行ったが, 会陰部皮膚の再発なども合併して初回手術後6か月で死亡した.組織学的には, アポクリン腺原発汗腺癌であったが, 今回の症例は汗腺癌がPaget病変を合併した貴重な症例と考えられた.また, 従来いわれているように肛門周囲難治性皮膚湿疹では, Paget病などの存在を十分考慮した診断が重要と考えられ, 積極的な外科的治療が必要と考えられた.