Biomedical subjects

H Yamazaki

Publications and source records attributed to H Yamazaki.

At least 181 records · Page 10Linked to original sources

PubMed1999

Fragile histidine triad transcription abnormalities and human papillomavirus E6-E7 mRNA expression in the development of cervical carcinoma.

BACKGROUND: Human papillomaviruses (HPV) are the most important etiologic factor for cervical carcinoma. However, additional cellular events may be necessary for malignant progression in the cervix. Recently, the fragile histidine triad (FHIT) gene, which is frequently lost in many cancers, was identified as a candidate tumor suppressor gene at chromosome 3p locus 14.2. METHODS: Thirty-five invasive cervical carcinomas (ICC), 13 Grade 2-3 cervical intraepithelial neoplasia specimens (high grade CIN), and 21 normal cervices were examined for expression of the FHIT gene and oncogenic HPV E6-E7 mRNA by reverse transcription-polymerase chain reaction (RT-PCR), and all of these FHIT cDNA were directly sequenced. RESULTS: Fourteen of 35 ICC (40%) showed abnormal FHIT transcription, whereas none of the high grade CIN or normal cervices had FHIT abnormalities. FHIT abnormalities included 13 short transcripts that were variously truncated between exons 4 and 8, 3 short transcripts with unknown sequences inserted, and 1 with no transcription. FHIT abnormalities were observed in all the small cell carcinomas, 4 of 7 adenocarcinomas and adenosquamous cell carcinomas (57%), and 7 of 25 squamous cell carcinomas (28%). Small cell carcinoma appeared to express the aberrant transcripts more frequently than squamous cell carcinoma (P=0.037). Immunohistochemical analysis showed that four ICCs with abnormal FHIT transcription did not express FHIT protein, whereas five of six CIN and ICC with no FHIT abnormality did express it. When expression of HPV E6-E7 mRNA was examined in the samples positive for HPV-DNA, E6-E7 transcripts were expressed in 7 of 9 high grade CIN (78%) and 18 of 28 ICC (64%). There was no difference in the frequency of HPV E6-E7 expression between CIN and ICC. In contrast, HPV E6-E7 expression was repressed in 8 of 10 (80%) ICC with FHIT abnormalities and in 2 of 18 (11%) in ICC with no FHIT abnormalities. CONCLUSIONS: Although E6-E7 genes of oncogenic HPV are important for the development of CIN and ICC, they may not be required for the development of carcinoma cells that have abnormal FHIT genes.

Acid Anhydride Hydrolases↗

PubMed1999

Perioperative fractionated high-dose rate brachytherapy for malignant bone and soft tissue tumors.

PURPOSE: To investigate the viability of perioperative fractionated HDR brachytherapy for malignant bone and soft tissue tumors, analyzing the influence of surgical margin. METHODS AND MATERIALS: From July 1992 through May 1996, 16 lesions of 14 patients with malignant bone and soft tissue tumors (3 liposarcomas, 3 MFHs, 2 malignant schwannomas, 2 chordomas, 1 osteosarcoma, 1 leiomyosarcoma, 1 epithelioid sarcoma, and 1 synovial sarcoma) were treated at the Osaka University Hospital. The patients' ages ranged from 14 to 72 years (median: 39 years). Treatment sites were the pelvis in 6 lesions, the upper limbs in 5, the neck in 4, and a lower limb in 1. The resection margins were classified as intracapsular in 5 lesions, marginal in 5, and wide in 6. Postoperative fractionated HDR brachytherapy was started on the 4th-13th day after surgery (median: 6th day). The total dose was 40-50 Gy/7-10 fr/4-7 day (bid) at 5 or 10 mm from the source. Follow-up periods were between 19 and 46 months (median: 30 months). RESULTS: Local control rates were 75% at 1 year and 48% in 2 years, and ultimate local control was achieved in 8 (50%) of 16 lesions. Of the 8 uncontrolled lesions, 5 (63%) had intracapsular (macroscopically positive) resection margins, and all the 8 controlled lesions (100%) had marginal (microscopically positive) or wide (negative) margins. Of the total, 3 patients died of both tumor and metastasis, 3 of metastasis alone, 1 of tumor alone, and 7 showed no evidence of disease. Peripheral nerve palsy was seen in one case after this procedure, but no infection or delayed wound healing caused by tubing or irradiation has occurred. CONCLUSION: Perioperative fractionated HDR brachytherapy is safe, well tolerated, and applicable to marginal or wide surgical margin cases.

Adolescent↗

PubMed1999

Highly sensitive high-performance liquid chromatographic assay for coumarin 7-hydroxylation and 7-ethoxycoumarin O-deethylation by human liver cytochrome P450 enzymes.

A highly sensitive method for the determination of coumarin 7-hydroxylation and 7-ethoxycoumarin O-deethylation by human cytochrome P450 (P450 or CYP) enzymes was developed using high-performance liquid chromatography (HPLC). The newly developed HPLC method was found to be about 100-fold more sensitive than the previous spectrofluorimetric method in detecting the metabolite 7-hydroxycoumarin (umbelliferone). With this high sensitivity, the kinetics of coumarin 7-hydroxylation and 7-ethoxycoumarin O-deethylation catalyzed by human liver microsomal and recombinant P450 enzymes were determined more precisely. With 36 different substrate concentrations in these two reactions, coumarin 7-hydroxylation was found to be catalyzed mainly by a single enzyme CYP2A6 and 7-ethoxycoumarin was oxidized by at least two enzymes CYP2E1 and CYP1A2 in human liver microsomes.

Chromatography, High Pressure Liquid↗

PubMed1999

Delayed visual NA potential in remitted schizophrenia: a new vulnerability marker for psychotic relapse under low-dose medication.

BACKGROUND: Lasting cognitive dysfunction throughout remission has been regarded as a biological vulnerability in schizophrenia, which may produce psychotic relapses with characteristic symptoms. Our hypothesis was that an abnormality in event-related potentials (ERPs) may be a neurophysiological marker of vulnerability to psychotic relapse in remitted schizophrenia. We conducted a 2-year follow-up study after evaluating ERP abnormalities to find a new ERP marker for schizophrenic relapse. METHODS: Visual ERPs were recorded from outpatients with remitted schizophrenia under maintenance pharmacotherapy (n = 44) and normal controls (n = 20) during a letter discrimination task. Based on the prospective study, the patients were divided into a relapse group (n = 20) and a nonrelapse group (n = 24). ERP findings that related to psychotic relapse within 2 years were analyzed. RESULTS: Compared with controls, the relapsers showed ERP abnormalities in the NA, N2, and P3 components, and the nonrelapsers in the P3 component. The peak latency of the NA potential was delayed significantly in the relapse group relative to the nonrelapse group, and predicted a psychotic relapse with about 90% probability. CONCLUSIONS: The delayed NA, which reflects early perceptual disorganization, may be a promising neurophysiological predictor of psychotic relapse in remitted schizophrenia under maintenance pharmacotherapy.

Adult↗

PubMed1999

Derivation of melanocytes from embryonic stem cells in culture.

We report that embryonic stem (ES) cells were efficiently induced to differentiate to melanocytes in vitro. When undifferentiated ES cells were cocultured with a bone marrow-derived stromal cell line, a very small but significant number of melanocytes were reproducibly generated. This process was greatly enhanced by addition of dexamethasone to the culture and strictly depended on steel factor, the ligand for the c-Kit receptor tyrosine kinase. Expression of c-Kit on the precursor cells was confirmed by using SCL/tal-1-/- ES cells, which are defective for producing hematopoietic cells, which were thus ruled out as possible sources of nonmelanogenic c-Kit-expressing cells. The morphology, reactivity to growth factors, and expression of melanogenic markers of the cells generated all indicated unequivocally that these cells were melanocytes. This culture system may provide a potent tool for the study of development and function of melanocytes.

Animals↗

PubMed1999

Increased interleukin-1beta production in the synovium of glenohumeral joints with anterior instability.

Macroscopic synovitis of the glenohumeral joint is frequently seen during arthroscopy in patients with anterior instability. Interleukin-1beta is known to be expressed in inflamed tissue, to correlate with the magnitude of inflammation, and to affect articular cartilage in the joint. We hypothesized that chronic synovitis may occur in the glenohumeral joint in patients with anterior instability. The purpose of this study was to examine the expression of interleukin-1beta in the synovium of the glenohumeral joint with anterior instability and to discuss its clinicopathologic significance. Specimens of synovial tissue around the greater tuberosity in the subacromial synovium (as controls) and around the rotator interval in the glenohumeral synovium were obtained from 10 patients who had anterior instability without signs of subacromial impingement. Semiquantitative reverse transcriptase-polymerase chain reaction was used to compare the levels of interleukin-1beta mRNA expression in the glenohumeral joint with those in the subacromial bursa. We also employed immunohistochemistry and in situ reverse transcriptase-polymerase chain reaction to detect the cells producing interleukin-1beta protein and mRNA. The levels of interleukin-1beta mRNA expression were significantly higher in the glenohumeral joint than in the subacromial bursa (p < 0.01). Histology showed nonspecific inflammation in all 10 samples of glenohumeral synovium, whereas no inflammation was seen in seven of 10 samples of subacromial synovium. Immunohistochemistry identified interleukin-1beta protein in the vessels and inflammatory and synovial cells (from lining to sublining layers) in synovium of the glenohumeral joint, whereas immunoreactivity was negative in seven subacromial bursa. The remaining three synovial specimens of subacromial bursa, however, showed positive immunoreactivity that was unremarkable and confined around the vessels. In situ reverse transcriptase-polymerase chain reaction was exclusively performed in the synovial specimens of the glenohumeral joint, which exhibited a positive reaction (in the same kinds of cells as seen with immunohistochemistry) in the lining and sublining layers and to a lesser extent in the stroma. Thus, our data confirmed the increased production of interleukin-1beta in the synovium of the glenohumeral joint in patients with anterior instability, suggesting the presence of chronic inflammation at the site. We conclude that this chronic synovitis may be partly associated with the development of dislocation arthropathy in the long term.

Acromion↗

PubMed1999

Roles of CYP2A6 and CYP2B6 in nicotine C-oxidation by human liver microsomes.

Nicotine C-oxidation by recombinant human cytochrome P450 (P450 or CYP) enzymes and by human liver microsomes was investigated using a convenient high-performance liquid chromatographic method. Experiments with recombinant human P450 enzymes in baculovirus systems, which co-express human nicotinamide adenine dinucleotide phosphate (reduced form) (NADPH)-P450 reductase, revealed that CYP2A6 had the highest nicotine C-oxidation activities followed by CYP2B6 and CYP2D6; the Km values by these three P450 enzymes were determined to be 11.0, 105, and 132 microM, respectively, and the Vmax values to be 11.0, 8.2, and 8.6 nmol/min per nmol P450, respectively. CYP2E1, 2C19, 1A2, 2C8, 3A4, 2C9, and 1A1 catalysed nicotine C-oxidation only at high (500 microM) substrate concentration. CYP1B1, 2C18, 3A5, and 4A11 had no measurable activities even at 500 microM nicotine. In liver microsomes of 16 human samples, nicotine C-oxidation activities were correlated with CYP2A6 contents at 10 microM substrate concentration, whereas such correlation coefficients were decreased when the substrate concentration was increased to 500 microM. Contribution of CYP2B6 (as well as CYP2A6) was demonstrated by experiments with the effects of orphenadrine (and also coumarin and anti-CYP2A6) on the nicotine C-oxidation activities by human liver microsomes at 500 microM nicotine. CYP2D6 was found to have minor roles since quinidine did not inhibit microsomal nicotine C-oxidation at both 10 and 500 microM substrate concentrations. These results support the view that CYP2A6 has major roles for nicotine C-oxidation at lower substrate concentration and both CYP2A6 and 2B6 play roles at higher substrate concentrations in human liver microsomes.

Animals↗

PubMed1999

Genetic transformation of a Rhizomucor pusillus mutant defective in asparagine-linked glycosylation: production of a milk-clotting enzyme in a less-glycosylated form.

Rhizomucor pusillus 1116R3 has a defect in alg2 encoding a mannosyltransferase in the asparagine (N)-linked oligosaccharide biosynthetic pathway and produces proteins in less-glycosylated forms. For development of a genetic transformation system for this zygomycete, an uracil auxotroph (mutant 1116U17) as the host strain was derived by ultraviolet (UV) mutagenesis as 5-fluoroorotic acid-resistant colonies and the orotidine-5'-monophosphate (OMP) decarboxylase (pyr4) gene as a selection marker was cloned from the wild-type strain R. pusillus F27 by the polymerase chain reaction with primers designed on the basis of the pyr4 sequences from other fungi. The amino acid sequence of R. pusillus Pyr4 deduced from the nucleotide sequence showed high homology with the OMP decarboxylases from various fungi. The pyr4 gene on pUC19 (plasmid pRPPyr4) was introduced into protoplasts of R. pusillus 1116U17 by polyethylene glycol-assisted transformation. Transformation under optimized conditions yielded 5 Ura+ transformants with 1 microgram pRPPyr4 DNA and 1 x 10(7) viable protoplasts. Southern blot analysis of the genomic DNA from the transformants showed that multiple copies of the pRPPyr4 sequence were integrated into the genome by homologous recombination at the pyr4 locus. For the purpose of production of a milk-clotting aspartic proteinase (MPP) in a less-glycosylated form, mpp from the wild-type strain was cloned in pRPPyr4 and introduced into protoplasts of R. pusillus 1116U17. Transformants obtained in this way contained multiple copies of mpp at the chromosomal mpp locus and produced MPP as a mixture of molecules having no sugar chains and Man0-1GlcNAc2 at the two N-linked glycosylation sites in an amount about 12 times larger than the parent strain. The transformation system for R. pusillus 1116U17 would be useful for production of proteins with truncated N-linked oligosaccharide chains.

Amino Acid Sequence↗

PubMed1999

Peroneal nerve palsy caused by intraneural ganglion.

A case of peroneal nerve palsy caused by an intraneural ganglion is presented. The cystic mass was located posterolateral to the lateral femoral condyle and extended along the common peroneal nerve distal to the origin of the peroneus longus muscle. The nerve was compressed in the narrow fibro-osseous tunnel against the fibula neck and the tight origin of the peroneus longus muscle. The nerve was decompressed by complete tumor excision and transection of the origin of the peroneus longus muscle. Full recovery of nerve function was obtained in 6 months.

Cysts↗

PubMed1999

Recurrence of medulloblastoma 19 years after the initial diagnosis.

A 25-year-old woman presented with recurrent medulloblastoma. She had been diagnosed with cerebellar medulloblastoma and treated for it at the age of 5 years. The new tumor was partially resected and chemotherapy was begun. However, a disseminated tumor of the spine was discovered and, despite radiotherapy, the patient became comatose and died. Histological examination of the tumor specimen showed characteristics similar to those of the first tumor. Late recurrence of medulloblastoma is rare, and a latency period of 19 years is the longest reported. This case violates Collins' rule and exceeds other proposed periods of risk. Extension of the period of follow-up in patients with medulloblastoma is advocated.

Adult↗

PubMed1999

CD9 molecule expressed on stromal cells is involved in osteoclastogenesis.

Osteoclasts are derived from hematopoietic stem cells and their development is dependent on the products of stromal cells. CD9, a member of the tetraspan transmembrane-superfamily, is expressed on both hematopoietic cells and stromal cells. Addition of antagonistic rat anti-mouse CD9 antibody (KMC8.8) to cultures inhibited osteoclastogenesis on established stromal cell layers. When rat bone marrow cells depleted of adherent stromal cells were cultured on mouse stromal cells, numerous tartrate-resistant acid phosphatase-positive multinuclear cells were observed, and KMC8.8, which recognizes mouse but not rat CD9, completely prevented the generation of osteoclasts, suggesting that the CD9 expressed on the stromal cell is essential for osteoclastogenesis. Possibly for the same reason, KMC8.8 pretreatment of the mouse macrophage-like cell line C7, which is able to differentiate into mature osteoclasts, did not inhibit subsequent C7 cell differentiation, whereas the addition of KMC8.8 to cocultures of C7 cells with stromal cells inhibited the differentiation of C7 cells into osteoclasts. Moreover, we found that blockage of a signal via CD9 on stromal cells reduced transcription of the osteoclast differentiation factor (Odf) gene, which, together with macrophage colony-stimulating factor, is essential for osteoclastogenesis. These results revealed that CD9 molecules on stromal cells play a critical role in osteoclast development, possibly by modulating the expression of Odf.

Animals↗

PubMed1999

Expression pattern of vascular endothelial growth factor isoform is closely correlated with tumour stage and vascularisation in renal cell carcinoma.

Vascular endothelial growth factor (VEGF) has five isoforms (VEGF206, 189, 165, 145 and 121). Increased VEGF expression in renal cell carcinoma (RCC) is associated with angiogenesis, but, it is not apparent which isoform is involved in this effect. We examined the isoform patterns of VEGF by reverse transcription-polymerase chain reaction (RT-PCR) in 47 RCCs. All showed increased VEGF expression as compared with extraneoplastic renal tissue. Four of the 47 RCCs showed VEGF121 alone, 10 showed VEGF121 + 165, and 33 showed the VEGF121 + 165 + 189 pattern. Patients with pathological stage pT3-4 RCC showed the VEGF121 + 165 + 189 isoform pattern at a significantly higher incidence (10/10, 100%) than those with pT0-2 (23/37, 62%) (P < 0.022). The VEGF121 + 165 + 189 isoform pattern was also significantly associated with high vessel counts and density (P = 0.0002, Mann-Whitney U test). These observations suggested that the VEGF189 mRNA isoform is closely associated with angiogenesis and results in the growth of RCC.

Blotting, Northern↗

PubMed1999

Thrombospondin-2 (TSP2) expression is inversely correlated with vascularity in glioma.

Thrombospondins (TSPs) are angiostatic factors in various cancers. However, the significance of TSPs has not been well characterised in glioma. We examined TSP1, TSP2 and vascular endothelial growth factor (VEGF) gene expression by reverse transcription-polymerase chain reaction (RT-PCR) in 37 gliomas. Thirty of the 37 glioma specimens showed VEGF gene expression. Eighteen of the 37 gliomas expressed the TSP1 gene. Seven gliomas lacked TSP2 gene expression, while the other 30 expressed TSP2. The lack of TSP2 gene expression was significantly associated with higher histological grade (Fisher's test, P = 0.0019) and increased vessel counts and density (Student's t-test, P < 0.0001), while there were no correlations between TSP1 and VEGF gene expression and clinicopathological features. These results indicate that the lack of TSP2 gene expression is a potent factor for enhancement of angiogenesis in glioma.

Adult↗

PubMed1999

Multidrug resistance-associated protein (MRP) expression is correlated with expression of aberrant p53 protein in colorectal cancer.

Multidrug resistance-associated protein (MRP) is one of the major factors responsible for non-P-glycoprotein (Pgp)-mediated multidrug resistance of human tumour cells. In this study, we examined MRP and aberrant p53 expression in 54 colorectal cancers (CRC), 35 carcinoma in adenomas (CIA) and 40 adenomatous polyps by immunohistochemical procedures. 38 of 54 (70%) CRCs, 16 of 35 (46%) CIAs and 3 of 40 (8%) adenomatous polyps were MRP positive (chi 2 test, P < 0.0001). 36/54 (67%) CRCs, 10/35 (29%) CIAs and 0/40 adenomatous polyps were p53 positive. 30 of the 36 p53-positive CRCs were also MRP positive and 8/10 CIAs were both p53 and MRP positive. MRP overexpression correlated with aberrant p53 accumulation in CRCs and CIAs (chi 2 test, P < = or 0.01). Coexpression of MRP and p53 in the same cells was confirmed in the CRCs and CIAs by double staining procedures. These results suggested that MRP overexpression is related to aberrant p53 expression in CRC.

ATP-Binding Cassette Transporters↗

PubMed1999

Cell-retained isoforms of vascular endothelial growth factor (VEGF) are correlated with poor prognosis in osteosarcoma.

Vascular endothelial growth factor (VEGF) is a major angiogenic factor. Osteosarcoma is characterised by hypervascularity and metastatic potential. We examined VEGF mRNA expression, VEGF isoform pattern and VEGF receptor (flt-1 and KDR) by RT-PCR analysis in 30 osteosarcomas. All 30 osteosarcomas expressed VEGF mRNA. 17 osteosarcomas (57%) expressed flt-1 mRNA, whilst 20 (67%) expressed KDR mRNA. 6/30 (20%) osteosarcomas were positive for VEGF121 only, 8 (27%) for VEGF121 + VEGF165, and 16 (53%) for VEGF121 + VEGF165 + VEGF189. Patients with osteosarcomas with VEGF165 (n = 24) had significantly poorer prognosis in comparison with those without VEGF165 (P = 0.022, Wilcoxon's test). The osteosarcomas with VEGF165 had significantly increased vascularity assessed on sections immunostained for CD34 (P < 0.001, Mann-Whitney U test). Although VEGF165 is a soluble isoform, it is also retained on the cellular surface. These results suggest that cell-retained VEGF isoforms (VEGF165, VEGF189) might be essential for neovascularisation in osteosarcoma, whilst the soluble VEGF121 isoform is not sufficient to stimulate neovascularisation in this type of neoplasm.

Adult↗

PubMed1999

Thrombospondin 2 expression is correlated with inhibition of angiogenesis and metastasis of colon cancer.

Two subtypes of thrombospondin (TSP-1 and TSP-2) have inhibitory roles in angiogenesis in vitro, although the biological significance of these TSP isoforms has not been determined in vivo. We examined TSP-1 and TSP-2 gene expression by reverse transcription polymerase chain reaction (RT-PCR) analysis in 61 colon cancers. Thirty-eight of these 61 colon cancers were positive for TSP-2 expression and showed hepatic metastasis at a significantly lower incidence than those without TSP-2 expression (P = 0.02). TSP-2 expression was significantly associated with M0 stage in these colon cancers (P = 0.03), whereas TSP-1 expression showed no apparent correlation with these factors. The colon cancer patients with TSP-2 expression showed a significantly low frequency of liver metastasis correlated with the cell-associated isoform of vascular endothelial growth factor (VEGF-189) (P = 0.0006). Vascularity was estimated by CD34 staining, and TSP-2(-)/VEGF-189(+) colon cancers showed significantly increased vessel counts and density in the stroma (P < 0.0001). TSP-2(-)/VEGF-189(+) colon cancer patients also showed significantly poorer prognosis compared with those with TSP-2(+)/VEGF-189(-) (P = 0.0014). These results suggest that colon cancer metastasis is critically determined by angiogenesis resulting from the balance between the angioinhibitory factor TSP-2 and angiogenic factor VEGF-189.

Analysis of Variance↗

PubMed1999

Administration of G-CSF to normal individuals diminishes L-selectin+ T cells in the peripheral blood that respond better to alloantigen stimulation than L-selectin- T cells.

To determine whether administration of G-CSF induces phenotypic or functional changes in T cells, we examined peripheral blood T cells from normal individuals receiving G-CSF for activation antigen and adhesion molecule expression before and after G-CSF administration. G-CSF (10 microg/kg/day) was administered subcutaneously to 14 normal individuals for 3-5 days and their PBMC were serially analyzed with monoclonal Ab (mAb) directed to HLA-DR, CD45RO, CD45RA, CD25, CD122, CD95, CD11a, CD49d, CD44 and CD62L (L-selectin) coupled with anti-CD3 mAb. Among T cells positive for these antigens, only the proportion of T cells expressing L-selectin significantly decreased from 68% to 37% after 3-day G-CSF administration. When peripheral blood CD3+ T cells obtained before and after G-CSF administration were sorted into two populations depending on the expression of L-selectin and tested for their proliferative response to allogeneic B cells, the reactivity of L-selectin- cells to alloantigen stimulation was consistently lower than that of L-selectin+ cells regardless of the exposure to G-CSF. The decrease in the relative number of L-selectin+ cells induced by G-CSF administration may contribute to the unexpectedly low incidence of severe acute GVHD after allogeneic PBSC transplantation.

Granulocyte Colony-Stimulating Factor↗

PubMed1999

High inducibility of heat shock protein 72 (hsp72) in peripheral blood mononuclear cells of aplastic anaemia patients: a reliable marker of immune-mediated aplastic anaemia responsive to cyclosporine therapy.

To better characterize immunologic aberrations in aplastic anaemia (AA), we examined peripheral blood mononuclear cells (PBMC) of 67 patients with AA and other patients with various haematological diseases for the expression of heat shock protein 72 (hsp72), which is inducible in lymphocytes by various stressors including antigenic stimulation. When freshly obtained PBMC were examined using flow cytometry, the proportion of cells expressing hsp72 in cytoplasm was significantly higher in allogeneic marrow transplant recipients (22 +/- 15%, mean +/-standard deviation (SD)) and AA patients (17 +/- 21%) than in normal individuals (6 +/- 3%). When PBMC were tested after heat treatment, only the proportion of hsp72+ cells of AA patients (37 +/- 30%) was significantly higher than that of the normal control (17 +/- 11%). Dual fluorescence analysis of the PBMC revealed that the majority of hsp72+ cells was CD3+. For 28 untreated AA patients, the proportion of hsp72+ cells in those who later responded to cyclosporine (CyA) (62 +/- 24%) was higher than that in non-responders (19 +/- 13%). Immunoblotting analysis revealed predominant expression of hsp72 in T cells. These findings indicate that high inducibility of hsp72 in PBMC by heat treatment is an immunologic aberration characteristic of CyA-responsive AA and that this simple test may be useful for identifying a subset of AA patients responsive to immunosuppressive therapy.

Adolescent↗