High-intensity focused ultrasound treatment for skin: ex vivo evaluation.

BACKGROUND/PURPOSE: High-intensity focused ultrasound (HIFU) has been used for skin tightening. However, there is a rising concern of irreversible adverse effects. Our aim was to evaluate the depth of thermal injury zone after HIFU energy passes through different condition. MATERIALS AND METHODS: To analyze the consistency of the HIFU device, phantom tests were performed. Simulations were performed on ex vivo porcine tissues to estimate the area of the thermal coagulation point (TCP) according to the applied energy and skin condition. The experiment was designed in three orientations: normal direction (from epidermis to fascia), reverse direction (from fascia to epidermis), and normal direction without epidermis. RESULTS: The TCP was larger and wider depending on the applied fluence and handpieces (HPs). When we measured TCP in different directions, the measured area in the normal direction was more superficially located than that in the reverse direction. The depth of the TCP in the porcine skin without epidermis was detected at 130% deeper than in skin with an intact epidermis. CONCLUSION: The affected area by HIFU is dependent on the skin condition and the characteristics of the HP and applied fluence. Considerations of these factors may be the key to minimize the unwanted adverse effects.

Comprehensive histologic analysis of interstitial lipolysis with the 1444 nm wavelength during a 3-month follow-up.

A number of near-infrared wavelengths have been proposed and studied for laser lipolysis, but the histologic evaluation of tissue response to laser lipolysis during long-term follow-up has been lacking. A 1444 nm Nd:YAG laser with better absorption in both fat and water has recently attracted attention. The present study was designed to investigate the comprehensive histopathology of 1444 nm Nd:YAG laser-assisted lipolysis at different energy levels during a 3-month follow-up. Laser lipolysis was performed on porcine fat tissue in vivo using a 1444 nm Nd:YAG laser (AccuSculpt®, Lutronic Corporation, Ilsan, Republic of Korea) and the total energies delivered interstitially to 10x10 cm² areas were 750 J, 1500 J, 2250 J, 3000 J, 3750 J, 4500 J, and 5250 J. Biopsy samples were taken and histologically analyzed immediately after biopsy and at 1, 2, 4, and 12 weeks postoperatively. With a fluence setting above 3000J/100 cm², inflammation was severe and remained by the 3-month follow-up, resulting in severe scarring of the fat tissue. Below this energy level, mild lobular inflammation in the early phase biopsy had resolved with no scarring by the 3-month follow-up. No histologic changes in the epidermis or dermal connective tissue were present. This study suggested that controlling the energy level is important for clinical applications of laser lipolysis with no significant complications.

MUC1 expression in human prostate cancer cell lines and primary tumors.

MUC1 expression was evaluated in normal prostate epithelial cells (PrEC), and prostate cancer cell lines in response to dihydrotestosterone (DHT), interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) treatment. Expression of MUC1 core protein was stimulated in PrEC and PC-3 cells after cytokine treatment, but was highly and constitutively expressed by DU-145 cells. MUC1 was not expressed by LNCaP, C4-2 or C4-2B cells under any condition. DHT alone or in combination with cytokines had no effect on MUC1 expression in any cell line tested. Using antibodies capable of detecting all isoforms of MUC1 core protein independent of their glycosylation state, immunohistochemical staining of tissue microarrays containing both nontumor and tumor tissue revealed that only 17% of tumor tissues and 41% of nontumor tissues stained positively for MUC1. Staining patterns in tumor tissue varied from focal apical staining to diffuse cytoplasmic staining. Neither the presence of MUC1 core protein nor its subcellular distribution correlated with Gleason grade. These data indicate that MUC1 is a poor marker of prostate cancer progression. Furthermore, IFN-gamma and TNF-alpha strongly induce MUC1 expression in both normal prostate epithelia and certain prostate tumor cell lines and may exacerbate pathologies associated with MUC1-positive prostate cancers.

Isolation and characterization of the major form of human MUC18 cDNA gene and correlation of MUC18 over-expression in prostate cancer cell lines and tissues with malignant progression.

Ectopical expression of huMUC18, a cell adhesion molecule in the immunoglobulin gene superfamily, causes a non-metastatic human melanoma cell line to become metastatic in a nude mouse system. To determine if MUC18 expression correlates with the development and malignant progression of prostate cancer, we investigated differential expression of human MUC18 (huMUC18) in normal prostate epithelial cells, prostate cancer cell lines, and prostatic normal and cancer tissues. We cloned and characterized the human MUC18 (huMUC18) cDNA gene from three human prostate cancer cell lines and three human melanoma cell lines. The cDNA sequences from the six human cancer cell lines were identical except differences in one to five nucleotides. The deduced amino acid sequences of the longest ORF were 646 amino acids that were identical in these cDNAs except for one to three amino acid residues. The amino acid sequences of all our huMUC18 cDNA genes are similar to that cloned by other group (GenBank access #M28882) except differences in the same seven amino acids. We conclude that huMUC18 cDNA gene reported here represents the gene product from a major allele. The MUC18 mRNA and protein was expressed in three metastatic prostate cancer cell lines (TSU-PR1, DU145, and PC-3), but not in one non-metastatic prostate cancer cell line (LNCaP.FGC). The expression of huMUC18 in these four cell lines is positively related to their extent of in vitro motility and invasiveness and in vivo metastasis in nude mice. HuMUC18 protein was also expressed at high levels in extracts prepared from tissue sample sections containing high grade prostatic intraepithelial neoplasia (PIN), but weakly expressed in extracts prepared from cultured primary normal prostatic epithelial cells and the normal prostate gland. Immunohistochemical analysis showed that huMUC18 was expressed at higher levels in the epithelial cells of high-grade PIN and prostatic carcinomas, and in cells of a perineural invasion, a lymph node, and a lung metastases compared to that in normal or benign hyperplastic epithelium (BPH). We therefore conclude that MUC18 expression is increased during prostate cancer initiation (high grade PIN) and progression to carcinoma, and in metastatic cell lines and metastatic carcinoma. Increased expression of MUC18 is implicated to play an important role in developing and malignant progression of human prostate cancer. Furthermore, the lacking of predominant cytoplasmic membrane expression of MUC18 appeared to correlate with malignant progression of prostate cancer.

Expression of a human cell adhesion molecule, MUC18, in prostate cancer cell lines and tissues.

BACKGROUND: Over expression of huMUC18, a cell adhesion molecule in the immunoglobulin gene superfamily, causes a non-metastatic human melanoma cell line to become metastatic in a nude mouse system. To determine if MUC18 expression correlates with the malignant progression of prostate cancer, we investigated differential expression of human MUC18 (huMUC18) in normal prostate epithelial cells, prostate cancer cell lines, and prostatic normal and cancer tissues. METHODS: RT-PCR and Western blot analyses were used to analyze the expression of MUC18 mRNA and protein in four human prostate cancer cell lines, cultured primary normal prostate epithelial cells, normal prostate and malignant prostate tissues. Immunohistochemistry was used to determine the expression of MUC18 antigen in prostatic tissues at different stages of malignancy. RESULTS: Human MUC18 mRNA and protein was expressed in three different prostate cancer cell lines (TSU-PR1, DU145, and PC-3), but not in one prostate cancer cell line (LNCaP.FGC). HuMUC18 protein was also expressed at high levels in extracts prepared from tissue sample sections containing high grade prostatic intraepithelial neoplasia (PIN), but weakly expressed in extracts prepared from either cultured primary normal prostatic epithelial cells or the normal prostate gland. Immunohistochemical analysis showed that huMUC18 was expressed at higher levels in the epithelial cells of high-grade PIN and prostatic carcinomas and in cells of a lymph node metastasis compared to that in normal or benign hyperplastic epithelium (BPH). CONCLUSIONS: We therefore conclude that MUC18 is expressed at higher levels in pre-malignant and malignant prostatic epithelium, including metastasis. We suggest that over-expression of MUC18 may be a new marker of human prostate cancer and also implicates its possible role in development and progression of prostate cancer.

Expression profiling of renal epithelial neoplasms: a method for tumor classification and discovery of diagnostic molecular markers.

The expression patterns of 7075 genes were analyzed in four conventional (clear cell) renal cell carcinomas (RCC), one chromophobe RCC, and two oncocytomas using cDNA microarrays. Expression profiles were compared among tumors using various clustering algorithms, thereby separating the tumors into two categories consistent with corresponding histopathological diagnoses. Specifically, conventional RCCs were distinguished from chromophobe RCC/oncocytomas based on large-scale gene expression patterns. Chromophobe RCC/oncocytomas displayed similar expression profiles, including genes involved with oxidative phosphorylation and genes expressed normally by distal nephron, consistent with the mitochondrion-rich morphology of these tumors and the theory that both lesions are related histogenetically to distal nephron epithelium. Conventional RCCs underexpressed mitochondrial and distal nephron genes, and were further distinguished from chromophobe RCC/oncocytomas by overexpression of vimentin and class II major histocompatibility complex-related molecules. Novel, tumor-specific expression of four genes-vimentin, class II major histocompatibility complex-associated invariant chain (CD74), parvalbumin, and galectin-3-was confirmed in an independent tumor series by immunohistochemistry. Vimentin was a sensitive, specific marker for conventional RCCs, and parvalbumin was detected primarily in chromophobe RCC/oncocytomas. In conclusion, histopathological subtypes of renal epithelial neoplasia were characterized by distinct patterns of gene expression. Expression patterns were useful for identifying novel molecular markers with potential diagnostic utility.

Immunohistochemical study of microphthalmia transcription factor and tyrosinase in angiomyolipoma of the kidney, renal cell carcinoma, and renal and retroperitoneal sarcomas: comparative evaluation with traditional diagnostic markers.

Angiomyolipoma has a unique immunophenotype with co-expression of muscle-specific actin and melanocytic markers such as HMB-45 and Melan-A. The most recently developed melanocytic markers, microphthalmia transcription factor and tyrosinase, have not been studied in the diagnosis of angiomyolipoma. We tested 29 renal angiomyolipomas (21 classic histology, 4 epithelioid variants, 2 lipomatous variants, and 2 leiomyomatous variants) with an immunohistochemical panel, including microphthalmia transcription factor, tyrosinase, HMB-45, Melan-A, and muscle-specific actin. Results were compared with 15 renal cell carcinomas (9 conventional types, 6 with sarcomatoid change), 2 leiomyosarcomas, 5 liposarcomas, and 1 unclassified high-grade sarcoma. Microphthalmia transcription factor expression was seen in 22 of 29 angiomyolipomas, one renal cell carcinoma, and one well-differentiated liposarcoma (that is, 2 of 23 non-angiomyolipomas; sensitivity 75%, specificity 91%). Tyrosinase expression was seen in 4 of 29 angiomyolipomas and 0 of 23 non-angiomyolipomas (sensitivity 14%, specificity 100%). HMB-45 was positive in 24 of 29 angiomyolipomas and 0 of 23 non-angiomyolipomas (sensitivity 83%, specificity 100%). Melan-A was expressed by 25 of 29 angiomyolipomas and 0 of 23 non-angiomyolipomas (sensitivity 86%, specificity 100%). Muscle-specific actin was expressed by 29 of 29 angiomyolipomas and 2 of 23 non-angiomyolipomas (both leiomyosarcomas; sensitivity 100%, specificity 91% [100% excluding leiomyosarcomas]). Microphthalmia transcription factor showed the most widespread staining in angiomyolipoma (50% of cases staining more than half of the tumor cells) followed by Melan-A (24% of cases staining more than 50%). Only three cases showed positivity for all four melanocytic markers, while in one case each only microphthalmia transcription factor and Melan-A were positive. We conclude that microphthalmia transcription factor, but not tyrosinase immunostaining, has a sensitivity and specificity that rivals those of the established markers, HMB-45 and Melan-A, in the diagnosis of angiomyolipoma. Our data supports the use of a panel in difficult cases that includes antibodies to microphthalmia transcription factor, either Melan-A or HMB-45, and muscle-specific actin to provide the best mix of high sensitivity, high specificity, nuclear and cytoplasmic immunolocalization, and widespread staining of cells within a given tumor.

Expression of progranulin and the epithelin/granulin precursor acrogranin correlates with neoplastic state in renal epithelium.

BACKGROUND: Current traditional pathological parameters, including staging and grading, are not sufficient in predicting outcome in patients with renal cell carcinoma (RCC). Acrogranin is an epithelial growth factor and has been demonstrated to play a role in teratocarcinogenesis and tumorigenesis. The aim of this study was to examine levels of acrogranin in renal cancer. MATERIALS AND METHODS: Western blot analysis was performed on renal tissue protein lysates. In addition, immunohistochemical (IHC) analysis of acrogranin expression was conducted on tissue sections of various histological types and grades of RCC. RESULTS: Western analysis showed that acrogranin levels were low in benign renal tissue and increased in malignant renal tissue. In addition, IHC revealed that high-grade RCC exhibited higher levels of expression than low-grade RCC and normal tissue. CONCLUSION: These data suggest that acrogranin may be a functional important growth factor in RCC and may be a potential molecular marker for high-grade RCC.

Methotrexate suppresses the interleukin-6 induced generation of reactive oxygen species in the synoviocytes of rheumatoid arthritis.

Various cytokines and reactive oxygen species (ROS) play a fundamental role in the inflammatory and immunologic processes of rheumatoid arthritis (RA). Methotrexate (MTX) is one of the disease-modifying anti-rheumatic drugs and its effect may be partly due to the modulation of immunologic or inflammatory reactions by some cytokines. In the present study, we investigated the effects of MTX on the gene expression and synthesis of interleukin-6 (IL-6), and the proliferative activity and the production of ROS in the fibroblast-like synoviocytes (FLSs) obtained from the patient of RA. The expression or production of IL-6 was induced spontaneously, and augmented by the addition of recombinant human IL-6 or recombinant human IL-1 beta and TNF-alpha in FLSs. These spontaneous and augmented IL-6 expressions or productions were suppressed by treatment with low-concentration of MTX (1 microg/ml). Also, IL-6 stimulated the proliferation of FLSs, and this IL-6 driven proliferation was inhibited with the treatment of MTX or N-acetylcysteine (NAC, 1 mM). Furthermore, ROS production in FLSs was increased significantly by IL-6, and its effect was also abrogated in the presence of MTX or NAC. These results suggest that inflammatory reaction in the synovium of RA patients could be augmented by the autocrine or other cytokine-induced production of IL-6 with subsequent generation of ROS in the synoviocytes, and the modulations of IL-6 synthesis and ROS production may contribute to the therapeutic effects of MTX for RA.

Relationship of cytokeratin 20 and CD44 protein expression with WHO/ISUP grade in pTa and pT1 papillary urothelial neoplasia.

The aim of this study was to assess the relationship of immunoreactivity of cytokeratin 20 (CK20) and CD44 across the spectrum of urothelial neoplasia using the WHO/ISUP consensus classification. A total of 120 papillary urothelial pTa and pT1 tumors (8 papillomas, 8 neoplasms of low malignant potential, and 42 low-grade and 62 high-grade carcinomas) were immunostained by using CK20 and CD44 antibodies. The relationships of tumor grade, pathologic stage, recurrences, and progression in stage with CK20 and CD44 immunoreactivity were assessed. WHO/ISUP grade correlated with tumor stage (P < 0.005), recurrence (P = 0.02), and progression in stage (P = 0.031). Normal urothelium showed CK20 immunoreactivity restricted to a few umbrella cells. Expression of CD44 in normal urothelium was restricted to the basal cell layer. Loss of CD44 immunoreactivity and increasing CK20 positivity were significantly associated with increasing tumor grade and stage (P < 0.005). An inverse relationship was observed in the staining patterns of CK20 and CD44 within individual cases, as well as in the aggregate data, with 79.2% of tumors with CD44 loss showing CK20 positivity (P < 0.001). In conclusion, CK20 and CD44 immunoreactivity are significantly related to the WHO/ISUP grade and to each other, and our data suggest their potential combined utility in predicting biologic behavior in patients with papillary urothelial pTa and pT1 neoplasms.

Congenital CD34-positive granular cell dendrocytosis.

Granular cell tumors involving the skin are mostly acquired lesions. The Schwann cell origin of these lesions is supported by positive immunostaining for S-100 protein and myelin basic protein. S-100- granular cell lesions rarely have been described in association with fibrous papules or dermatofibromas. The congenital variety of S-100- granular cell tumors occurs almost exclusively in the gingiva. The cell origin of these lesions is not well delineated. We report a hitherto undescribed case of a congenital cutaneous lesion which is histologically characterized by diffuse dermal infiltrates of S-100- but CD34+ granular dermal dendrocytes. The granular appearance of these CD34+ dendrocytes is attributed to an abundance of phagolysosomes. The pathogenetic mechanism of this unusual lesion remains to be elucidated.

Fetus-in-fetu in the scrotal sac of a newborn infant: imaging, surgical and pathological findings.

We report a case of fetus-in-fetu located in the scrotal sac of a newborn male infant. Plain radiography (including specimen radiography), ultrasonography and MRI clearly demonstrated vertebral column, ribs, skull, pelvic bones, femurs and a portion of tibiae and humeri. The diagnosis was confirmed by pathological examination.

Tissue transglutaminase is not increased during apoptosis of HT-1080 human fibrosarcoma cells.

Tissue transglutaminase (tTGase), a cytosolic enzyme which catalyzes the covalent cross-linking of proteins, is thought to be involved in the apoptosis. Here, we tested whether tTGase is involved during HT-1080 fibrosarcoma cell apoptosis induced by the YIGSR (Tyr-Ile-Gly-Ser-Arg) peptide. This sequence is derived from the laminin alpha1 chain, and its potency is increased by the formation of a 16mer polymerization using a lysine tree structure. Cells were treated with several different concentrations of Ac-Y 16 for 16 hours, and apoptosis was increased in dose-dependent manner. When assayed by incorporation of [14C] putrescine into succinylated casein, total transglutaminase activity was decreased in parallel with the change in the number of attached cells. Western blot analysis using polyclonal antibody against tTGase showed that the tTGase protein level had not been significantly changed when equal amounts of the protein were applied. To confirm this result, we induced apoptosis of these cells by coating the tissue culture plates with non-adhesive poly-hydroxyethyl methacrylate (HEMA). Western blot analysis showed that the tTGase protein level did not change during this process of apoptosis. Although it has been suggested that tTGase is involved in the process of apoptosis of various cells in vitro and in vivo, our data demonstrate that tTGase is not involved in the process of apoptosis of HT-1080 human fibrosarcoma cell induced by either Ac-Y 16 or a non-adhesive culture surface.

Mansonian schistosomiasis in rectum--report of a case.

Schistosomiasis is a snail-transmitted trematodiasis acquired by immersion in water which contains the cercariae. In Korea, six imported cases of urinary schistosomiasis by Schistosoma haematobium and one case of imported cerebral schistosomiasis by S. mansoni were reported. Herein we report a case of S. mansoni infecting rectum of a 46 year-old Korean male, who had been to Saudi Arabia for two years. On colonoscopy for routine physical check up, a 0.4 cm polyp in the rectum was detected and biopsy was done. Microscopically, rectal mucosa showed several granulomas which were composed of macrophages, lymphocytes, neutrophils and eosinophils. The center of each granuloma showed an ovoid egg often containing miracidium. The eggs measured 130 x 60 microns in average size. They had yellowish-brown transparent shell with the characteristic lateral spine. This is the 8th imported case of schistosomiasis in Korea and the second one infected by S. mansoni.

The significance of glomerular hypertrophy in focal segmental glomerulosclerosis.

It is not clear whether glomerular hypertrophy is linked to the pathogenesis of human focal segmental glomerulosclerosis (FSGS). To probe the significance of glomerular hypertrophy in relation to development of FSGS, we studied 16 adults with primary FSGS by morphometry, and the findings were compared to age- and sex-matched subjects with minimal lesion. Mean glomerular volume (MGV), mesangial volume density, mesangial volume per glomerulus, and cortical interstitial volume density [Vv(int/cortex)] were significantly increased in the FSGS patients when compared to the minimal lesion patients. The increase in mesangial volume in FSGS was mainly due to expansion of mesangial matrix. In FSGS, MGV was related directly to % of glomeruli with glomerulosclerosis (r = 0.47, p < 0.05), to mesangial volume per glomerulus (r = 0.57, p < 0.01), and to Vv(int/cortex) (r = 0.47, p < 0.05). The percentage of glomerulosclerosis correlated directly with Vv(int/cortex) (r = 0.83, p < 0.0005), and with mesangial volume per glomerulus (r = 0.47; p < 0.05) in FSGS. Also, there was a direct relationship between Vv(int/cortex) and mesangial volume per glomerulus (r = 0.49; p < 0.05) in FSGS. Glomerular hypertrophy observed in our patients with primary FSGS was intercorrelated with the extent of glomerulosclerosis, mesangial expansion and interstitial fibrosis. Glomerular hypertrophy seems to be one of the morphological facets present in FSGS. Glomerular hypertrophy tends to coexist with FSGS rather than precede its development. Thus, in biopsies diagnosed with minimal lesion the presence of glomerular hypertrophy appears to be an indication that the coexistent FSGS lesions are undetected as a result of sampling problems.

Curvilinear-gradient high-performance liquid chromatography for identification of mycobacteria.

Over a 1-year period, 502 mycobacterial cultures submitted to the Microbial Diseases Laboratory were identified by high-performance liquid chromatography (HPLC) in parallel with standard biochemical methods. Identification by HPLC using a curvilinear gradient was achieved by comparing the chromatograms of the unknown cultures to chromatograms for known reference strains, together with calculation of peak height or peak area ratios, as necessary. The overall agreement between HPLC and biochemical identification was 97.2%. In addition, 7 of 12 cultures of Mycobacterium bovis were identified by HPLC as the BCG strain. Of 111 cultures biochemically identified as members of the M. avium complex (MAC), 108 were confirmed as MAC by DNA probe and 106 were confirmed by HPLC. Of the latter 106, 58 probe-positive strains were identified as M. avium, 38 were identified as M. intracellulare, and 10 were identified as Mycobacterium sp. strain "X" by HPLC. Of the remaining five nonchromogenic cultures, four had MAC-like chromatograms that did not match any in our library sufficiently to permit definitive identification. Of the latter four, two were confirmed as MAC strains by DNA probe and two were not. The last of the cultures biochemically identified as MAC (1 of 111) was a mixture of MAC and non-MAC strains. Overall, only 2 of 502 cultures yielded results by HPLC that differed from those obtained by standard biochemical methods. The HPLC result was confirmed in both cases by an independent national reference laboratory. In the 12 instances in which HPLC did not provide identification, the chromatograms were either uninterpretable or did not match available reference chromatograms. These findings show that the identification obtained by HPLC concurs well with that obtained by both the standard biochemical methods and the DNA probes. Thus, identification by HPLC provides mycobacteriology laboratories with a reproducible and specific method for accurate and timely identification of most medically important mycobacteria.

Genotypic identification of pathogenic Mycobacterium species by using a nonradioactive oligonucleotide probe.

Commercial DNA hybridization assays (Syngene, Inc., San Diego, Calif.) utilizing alkaline phosphatase-labeled oligonucleotide probes for the identification of Mycobacterium tuberculosis complex and M. avium complex (MAC) were evaluated with 261 isolates of mycobacteria. On the basis of biochemical criteria, the test for MAC was 98% specific and more sensitive (95 of 99, 95%) than Gen-Probe (88 of 99, 89% sensitivity); the major difference in sensitivity noted between the two systems was related to the hybridization of seven MAC strains to the SNAP X probe. The M. tuberculosis complex probe correctly identified all 62 isolates of M. tuberculosis and all 11 isolates of M. bovis, for a sensitivity of 100%. There were two discrepant reactions with mycobacteria other than M. tuberculosis complex isolates.

Abnormalities of T-cell subsets in Behçet's syndrome.

T cells and T-cell subsets were determined in the peripheral blood of 12 patients with Behçet's syndrome and 30 normal healthy control subjects. When compared with the control group, the mean percentage of T cells for the group with Behçet's syndrome was significantly decreased (73% v 61%). The mean percentage of T mu (helper) cells for the group with Behçet's syndrome (26%) was also significantly decreased from the mean value of the control group (42%). There was a concomitant significant increase of T gamma (suppressor) cells of the group with Behçet's syndrome (19%) over the mean value of the control group (10%). These results clearly indicated that there were alterations of T cells and T-cell subsets in this disease.

Leprosy. XII. T-cell subsets in lepromatous leprosy.

The authors quantitated T-rosette-forming cell (TRFC) and T-cell subsets (T mu, T gamma) in the peripheral blood of twenty patients with lepromatous leprosy. The results obtained in their studies are as follows: (1) They reconfirmed the low levels of TRFC in patients with lepromatous type of leprosy; (2) T-cell subsets, both T mu (helper) and T gamma (suppressor) cells, showed lower levels in all patients with lepromatous leprosy than mean values of normal healthy controls; (3) The degree of decreased levels of T mu cells (96%) was more severe than other parameters TRFC (70%) and T gamma cells (47%) in all patients with lepromatous leprosy; and (4) It may be concluded that the alteration of the T-cell subset, T mu-cells, reflects a more fundamental abnormality than TRFC aberration in demonstrating the impairment of cell-mediated immunity in patients with lepromatous leprosy.

Leprosy XII. Quantitative analysis of thymus-derived lymphocyte response to phytohemagglutinin in leprosy.

The immune status of various leprosy patients was evaluated by using a micromethod to evaluate lymphocyte responses to phytohemagglutinin (PHA). In our study, whole blood was used and the degree of response to PHA stimulation was expressed in terms of unit volume of blood. A markedly decreased response to PHA stimulation was noted in patients with active lepromatous leprosy. Patients with active lepromatous leprosy who have been proved drug (DDS) resistant showed less response than did those of drug sensitive patients with active lepromatous disease, while the patients with active lepromatous leprosy complicated by erythema nodosum leprosum (ENL) showed higher response than did those of patients with no complicated ENL. Comparing the results obtained to those obtained using other methods for T cell analysis indicates that these results reflect the number of T lymphocytes in the leprosy patient. Thus, this simple method is of value in assaying the presence and responses of T lymphocytes in the leprosy patient.