Male New Zealand Black/KN mice: a novel model for autoimmune-induced permanent alopecia?

BACKGROUND: Irreversible, permanent and scarring alopecia is associated with several autoimmune diseases, including all autoimmune connective tissue disorders. The pathogenesis of autoimmune-induced permanent alopecia (APA) is still poorly understood, and instructive, simple mouse models for the study of APA are needed urgently. During the course of our studies in a well-established mouse model for chronic rheumatoid arthritis, the New Zealand Black/KN (NZB/KN) mouse, we noticed that ageing male NZB/KN mice developed spontaneous APA. OBJECTIVES: To study whether alopecia seen in ageing male NZB/KN mice displays key features of human APA and may, thus, be a useful new mouse model for clinically relevant APA research. METHODS: NZB/KN, the F1 hybrid of NZW/N Slc x NZB/KN (W/BKN F1), the F1 hybrid of NZB/KN x NZW/N Slc (BKN/W F1), and the F2 hybrid of W/BKN F1 x W/BKN F1 mice were employed in this study, in order to check which strain carries the highest risk of alopecia development. Besides routine histology, CD3, CD4 and CD8 expression as well as immunoglobulin (Ig) G and IgM deposition in hair follicles were investigated by immunohistology/immunofluorescence. Mast cell distribution/degranulation and Ki-67 (proliferation)/TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling) (apoptosis) positive cells were also analysed. RESULTS: Only F2 male NZB/KN mice were prone to develop alopecia, suggesting that Y chromosome-associated gene(s) are involved in the pathogenesis of APA, which incidence rises with increasing age. The lesional alopecia skin in 12-month-old male NZB/KN mice showed a sharp decline in hair follicle density, thus meeting a key criterion of permanent alopecia. Both macroscopically and histologically, the alopecia seen in these mice resembled in many respects different stages of clinical APA, such as alopecia associated with chronic discoid lupus erythematosus (DLE) in humans. Lesional APA hair follicles in mice displayed intrafollicular and perifollicular mononuclear cell infiltrates, as well as an increased number of activated (degranulated) perifollicular mast cells. In the fully developed lesion, many CD4+ cells were seen in perifollicular locations, including the epithelial stem cell region (bulge), and also contained a few CD8+ T cells. IgM deposits were found in the follicular basement membrane zone (BMZ). Both in the bulge and the hair matrix region of the affected anagen hair follicles, there were signs of massive keratinocyte apoptosis. CONCLUSIONS: Our currently available data suggest that male but not female NZB/KN mice may indeed represent a suitable mouse model for APA, with some similarities to the permanent alopecia seen in human DLE patients, although additional and confirmatory investigations are needed before this mouse strain can be accepted as a murine equivalent of APA in humans.

Susceptibility of T cell receptor-alpha chain knock-out mice to ultraviolet B light and fluorouracil: a novel model for drug-induced cutaneous lupus erythematosus.

The anticancer agent 5-fluorouracil (FU) frequently induces cutaneous lupus erythematosus (LE) lesions on sun exposed sites. Based on this observation, we have tried to establish a cutaneous LE model of C57BL/6 J (B6) mice, B6 T cell receptor (TCR)-alpha(-/-) mice and B6 TCR-delta(-/-) mice treated with FU and/or ultraviolet B light (UVBL) in order to clarify the role of T cells and the cytokine profile of cutaneous lupus lesions. Cutaneous LE-like skin lesions could be induced in TCR-alpha(-/-) mice with low FU (0.2 mg) plus UVBL, and in B6 mice treated with a high dose of FU (2.0 mg) plus UVBL. In contrast, low FU plus UVBL induced such skin lesions in TCR-delta(-/-) mice at a very low incidence. Specifically, the skin lesions of TCR-alpha(-/-) mice with low FU plus UVBL appeared more rapidly and were more severe than lesions in B6 mice. The former had the common characteristic features of human chronic cutaneous LE such as typical histology, positive IgG at the dermoepidermal junction, low antinuclear antibody and low mortality. Furthermore, a Th1 response was induced in the development of drug-induced cutaneous LE. FU and UVBL-induced cutaneous LE-like eruption is an excellent model for better understanding the pathomechanisms of skin lesion development in LE.

Slow acetylator genotypes as a possible risk factor for infectious mononucleosis-like syndrome induced by salazosulfapyridine.

We report two patients with infectious mononucleosis-like syndrome induced by salazosulfapyridine (SASP). In both cases, high fever, skin rash, liver dysfunction and atypical lymphocytosis developed 3 weeks after initiating treatment with SASP. SASP is known to be mainly metabolized by N-acetyltransferase 2 (NAT2), and acetylation phenotypes (rapid, intermediate and slow acetylator) correlate with NAT2* genotypes. In our two patients, we investigated NAT2* genotypes by the polymerase chain reaction-restriction fragment length polymorphism method. We identified NAT2*6/*7 in one patient, and NAT2*6/*5 in the other, suggesting that both were slow acetylator phenotypes. In 20 healthy volunteers we found no slow acetylator genotypes. Genotyping prior to medication may be useful in evaluating patients with a high risk of severe systemic reaction to SASP.

Discoid lupus erythematosus (DLE)-like lesion induced by uracil-tegafur (UFT).

A mixture of uracil and tegafur (UFT) is a common antineoplastic agent in Japan. We report a 64-year-old Japanese woman with discoid lupus erythematosus (DLE)-like lesions which were induced by UFT. After surgery to treat lung cancer, UFT (300 mg/day) was administered and she developed round erythema on her right cheek. A skin biopsy specimen taken from the right cheek site revealed atrophy of the epidermis, a slight liquefaction of the basal cell layer, and patchy lymphocytic infiltration in the perivascular and perifollicular regions. A test for antinuclear antibody was weakly positive (80 fold), and rheumatoid factor was slightly elevated (7.6 IU/ml). After discontinuation of UFT, the erythema completely regressed within 2 months. We reviewed 17 cases of DLE-like lesions induced by fluorouracil agents and summarized the common features.

PCNA and p53 in urinary bladder cancer: correlation with histological findings and prognosis.

BACKGROUND: This study aimed to immunohistochemically examine the expression of proliferating cell nuclear antigens (PCNA) and p53 protein in transitional cell carcinomas (TCC) of the urinary bladder, and to investigate possible correlations of this expression with the tumor grade or stage, tumor recurrence, and prognosis of the disease. MATERIALS AND METHODS: The immunohistochemical status of the PCNA and p53 proteins were determined on paraffin-embedded sections from 128 patients with TCC of the urinary bladder, using PC-10 and DO7 as the primary antibodies. Positive stainings were represented by scores (Labeling Index; LI, %) calculated as the percent of positive cells among all neoplastic cells counted. The findings were compared to the patients' histopathological features. Patients who underwent transurethral resection were divided into groups with "low" and "high" scores for PCNA and p53, respectively, and the tumor recurrence rate was compared among the groups. Patients who underwent total cystectomy were similarly divided into "low" and "high" score groups, and survival rates of the groups were compared. RESULTS: PCNA expression was observed in 66 of 128 patients (51.6%), with a mean labeling index of 26.6%. Overexpression of p53 was observed in 65 of 128 patients (50.8%), with a mean labeling index of 35.3%. There were significant correlations of the PCNA and p53 indices with both histological grade and stage of the tumor. In the TUR group, there were no statistically significant differences in recurrence rate between the groups with high or low scores for either PCNA or p53. In the total cystectomy group, there was a significant correlation between survival rate and positive staining for PCNA, but not for p53. The relationship between 2 parameters, PCNA and p53 scores, was not significant (linear correlation coefficient, r = 0.67). CONCLUSIONS: PCNA and p53 status in transitional cell carcinomas of the urinary bladder is related to the histopathological findings. We also suggest that immunohistochemical staining for PCNA provides significant clinical information which may be useful in the initial selection of therapy. However, overexpression of p53 does not appear to represent an independent prognostic marker in bladder tumors.

[Combined hormone and chemotherapy for the patients with advanced prostate cancer].

From November 1984 to December 1993, 72 patients with newly diagnosed, stage C or D prostate cancer were treated with chemohormonal therapy consisting of CDDP, CPM, diethylstilbestrol diphosphate and orchiectomy. Forty-four of 72 patients (61%) had partial response 3 months after the initiation of the treatment. A good response rate was observed in prostate gland and tumor markers (63% and 81%), but the response rate was poor in bone and other soft tissure metastasis (17% and 18%). Subjective symptoms improved in 82% of the pain, 70% of the infravesical obstruction, 65% of the performance status, and 5% of the body weight. The 5-year survival rate was 66%. The prognosis of the patients with poorly differentiated carcinoma or high Gleason score tumor was poor. Survival rate was no different between stage C and D. Relapse occurred in 16 patients (22%) and tended to occur in the patients with poorly differentiated carcinoma, high Gleason score tumor, stage D2 or stable disease. The survival rate was better, and the relapse rate lower than in the 35 patients with hormonal treatment alone during the same period, but no significant difference was observed between the two groups.

[Intraarterial chemotherapy for patients with invasive bladder cancer--clinical effects and preliminary results in bladder preservation].

Preoperative intraarterial chemotherapy including cisplatin, etoposide and adriamycin was performed for 16 patients with invasive bladder cancer between April, 1989 and September, 1993. Of 16 patients, radical cystectomy was done in 12 patients and the bladder was preserved by a transurethral resection in 4. Of 12 patients treated with cystectomy, complete response was achieved in 2 cases and partial response in 4 (response rate: 50%). Pathological evaluation of resected specimens revealed significant downstage of the tumors in 8 of 12 patients (67%). At follow-up 4 patients had recurrent and/or distant metastasis. Three of these four patients died of cancer progression and one survived with cancer. Of 4 patients with preserved bladder, complete response was achieved in one case and partial response in 3. Pathological specimens evaluated by transurethral resection showed significant downstage of tumors in all cases. One patient had an invasive, recurrent tumor in the preserved bladder and was treated with radical cystectomy 6 months after initiation of the therapy. All patients tolerated the treatment well. Intraarterial chemotherapy seems to be effective for invasive bladder cancer, but the true success of bladder-preserving treatment by a transurethral resection, chemotherapy and other therapy will require further study.

[Effective chemoembolization for liver metastasis in a case of testicular cancer].

A 21-year-old male with a previous history of retroperitoneal tumor treated with chemotherapy, radiotherapy, immunotherapy and probe laparotomy was admitted with a complaint of right scrotal enlargement and liver tumor. He had retroperitoneal mass and large liver tumor on an abdominal CT scan. Right inguinal orchiectomy was performed on April 14, 1988. Histopathological diagnosis confirmed embryonal cell carcinoma plus seminoma. Cisplatin, vinblastine and bleomycin combination chemotherapy (PEB) was started and repeated for 4 courses. Repeat CT scan showed significant shrinkage of retroperitoneal and liver tumor, but not complete eradication of tumors. Chemoembolization of liver tumor was done on August 5, 1988. Biopsy of liver tumor revealed disappearance of viable tumor cells. On CT scan, no retroperitoneal tumor was detected on February 27, 1990, and no liver tumor was delineated on August 30, 1991. The patient was alive with no evidence of disease on February 28, 1994.