For diagnosis of Legionnaires'disease, we developed a radioimmunoassay of urinary soluble antigen of L. pneumophila, Philadelphia 1.The bacterial solution was desinfected by heating and a soluble component was used as an antigen.Antiserum was obtained by innoculating rabbits with formalin-fixed bacteria and purified by salting out in ammonium sulfate and column chromatographies on DEAE cellulose and a Sephadex G-200.
Radioimmunoassay was carried out as follows: 1) incubate a 500μl urine sample or a standard soluble antigen with one peace of antibody-coated bead at 37°C for 90 min.2) further incubation with 350μ1 of ¹²⁵ I-labelled antibody at 37°C for 90 min.3) Counting radioactivities of 236 I with a γ-counter after washing a bead.The sensitivity of the assay was 12.5 unit/ml and the C.V.for each concentration of the standard curve was 2.7-9.2%.
The dilution test showed good result.As to a crossreactivities, the antibody showed a specific binding only to Serogroup I.The time course of urinary antigen was analyzed by radioimmunoassay after administering guinea pigs with 10⁹ cells/ml of L.pneumophila, Philadelphia 1.i.p., and it was initially detected in 8 out of 9 animals on the second day and concecutively until the third and eighth day.The remaining one was a case of negative reaction.
The soluble antigen was a macromolecule which was eluted at the void volume by gelfiltration with Sephadex G-200 or G-75, while the urine of the infected guinea pigs contained the antigen with various molecular weights.
The above results suggest that a radioimmunoassay of the soluble antigen is a useful routine method for a diagnosis of Legionnaires'disease because of its simple and timesaving features and also its capability of detecting the infection in the early stage.

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In our previous report, we presented the results of basical studies of detection of urinary Legionell antigen by radioimmunoassay for the diagnosis of Legionnaires'disease.
We could have diagnosed one sporadic case of Legionnaires'disease of a young male by using this method clinically.
This method is very available for the clinical diagnosis of this disease, for the specimens of the early stage or after the administration of effective antibiotics could be detected positive by this method.
And because of its superior sensitivity, we may be able to diagnose some cases which serum Legionella antibody titer is in upper limit of normal range.This is another availability of this method.
We presented the first case of Legionnaires' disease in Japan diagnosed by detection of urinary Legionella antigen by RIA.

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A ratio leukocidin-producing strains to clinical isolates was investigated together with production of protease, elastase and exotoxin.It was also examined whether these strains contain the common proteineous antigen (OEP) which resides in the cell wall.By the agar gel precipitin test using crystalline or highly purified antigens, leukocidin, protease, elastase, exotoxin and OEP, and their specific antisera, it was proved that leukocidin reacted only against the homologous serum.Similarly, the serological specificity of the other four antigens was also confirmed.
In order to detect production of leukocidin a strain to be tested was inoculated on one side of the Petri dish which was incubated at 37°C for 2 days and exposed to chloroform vapour.Then 5 wells were made at a distance of 5 mm from the growing culture, and these were filled with the 5 kinds of antisera. Precipitin reaction was judged after the dish was left at 25°C overnight.Consequently, 87 clinical isolates (96.7%) out of 90 were found to produce leukocidine.Protease, elastase and exotoxin were also prodused in high percentages.OEP was found to exist in all strains.

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In order to estimate anti-leukocidin antibody in sera of patients, an ELISA test using horseradish peroxidase conjugated protein A was devised.Sera of 39 cases of diffuse panbronchiolitis (DPB) were investigated for anti-leukocidin antibody.Mean antibody anti-leukocidin titer in sera of 17 DPB cases which had not been infected with Pseudomonas aeruginosa and 5 DPB cases which were transiently infected with the bacteria, were not significantly different from the mean anti-leukocidin titer in sera of 11 healthy control cases, while the mean anti-leukocidin titer in sera of 17 DPB cases persistently colonized was significantly higher than in the former cases.
These results indicate that leukocidin was produced at the local site of infection in DPB patients. Studies to clrify the role of leukocidin in the pathogenicity of DPB are needed.

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Kyushu is known as area in which adult T-cell leukemia (ATL) is endemic.In order to determine the distribution of antibody to ATL-associated antigen (anti-ATLA) in the Nichinan district of southern Kyushu, determination of anti-ATLA status of inpatients in a general hospital in Nichinan City, Miyazaki prefecture was carried out from August to December, 1983. Sera from 914 inpatients were tested for presence of anti-ATLA by Enzymeimmunoassay (EIA) prepared by Eisai Co., Ltd., Tokyo, Japan.
Results obtained are as follows
1) Overall prevalence of anti-ATLA was 18.9 per cent (173 of 914 individuals).Prevalence of anti-ATLA increased with age, reaching a maximum of 37.1 per cent for people 70 years old or over.
2) Prevalence of anti-ATLA was 18.1 per cent (73 of 403) in males and 19.6 per cent (100 fof 511) in females.Significant difference by sex was not recognized.
3) Anti-ATLA was most prevalent in patients with neoplasms (36.7%).Patients with following pathological conditions were included in this category (anti-ATLA prevalencies are given in parenthesis): malignant lymphoma (100%), leukemia (0%), stomach cancer (41.7%), and miscellaneous cancer (26.8%).After neoplasms, anti-ATLA was most prevalent in patients with diseases of the musculoskeletal system and connective tissue (33.3%) and diseases of the circulatory system (27.9%).
4) Mean optical density (OD) values by EIA were 1.034±1.035 for the 173 anti-ATLA-positive individuals and 0.038±0.023 for the 741 negative individuals;The difference between the values for the positive and negative sera was large.Furthermore, the mean OD value was significantly higher for anti-ATLA-positive males (1.142±0.888) than for anti-ATLA-positive females (0.955±1.180) (p<0.001).

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For the purpose of studying the pathogenesis of Japanese encephalitis, mice were inoculated with varied concentrations of Japanese encephalitis (JE) virus, Kamiyama strain, into the food pad of each mouse and treated with cyclophosphamide (CY) on the different day (s) before or after the virus inoculation.
The results obtained are as follows.
The administration of one dose of CY between two days before and four days after the virus inoculation increased the mortality of mice infected with JE virus.
The administrations of two or three doses of CY increased the mortality of mice infected with JE virus and the infectivity of mice inoculated with the virus. Particularly, in the groups treated with second inoculation of CY between five days and ten days after the virus inoculation mortality was increasedmarkedly.
Some of the JE virus infected mice treated with CY died of Japanese encephalitis more than twenty days after the virus inoculation.

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Protective activities of immnuoglobulin preparations depend on their contents of the specific antibodies against causative agents and are measured by the agglutination and neutralization titers.But human immunoglobulin preparations for intravenous injection are treated to remove small aggrgates in Cohn's fraction II preparation of Gammaglobulin.Because of these treatments, their agglutination and neutralization titers are not always parallel with their protective activities.
We examined antibody activities of sulfonated immunoglobulin (Venillon) by the binding activities with representative strains of possible opportunistic pathogenic species using ELISA technique instead of the agglutination test, and found that antibody activities did not decrease by the sulfonation. We also tested complement dependent bacteriolytic function of the sulfonated immunoglobulin by the survival rates and electronmicroscopic observation of lysed cells, and found that sulfonated immunoglobulin caused full bacteriolysis within 45 min.This result suggested that conformational changes of immunoglobulin by sulfonation reduced small aggregation which caused anaphylaxic adverse reactions but kept specific complement activation activity.
Through this estimation of antibody activities of the sulfonated immunoglobulin preparations, high neutralization titers against varicella virus, Staphylococcus epidermidis, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Enterobacter were confirmed.This might suggest the high protective activity of this sulfonated immunoglobulin preparation against the infections by these pathogens.

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Five strains of Clostridium difficile were isolated from specimens other than stool. One of these was isolated from a case of intraabdominal abscess after colectomy, two from decubital ulcers, one from a bile, and one from a blood.
All five cases were treated with antibiotics prior to recovery of C. difficile, and five strains of C. difficile were isolated with other aerobes and anaerobes concomitantly.

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[in Japanese]

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It is known that herpes zoster in provoked by the reactivation of varicella zoster virus which has been latent within the dorsal ganglion in immunosuppressed patient. Ara-A and acyclovir is used as antiherpes virus agent with insufficient effect in compromized hosts with severe generalized herpes zoster.
Recently, we administered cimetidine to a patient with herpes zoster which developed in the course of pneumonia with a marked effect. She was a 81-year-old woman with no malignancy. Severe pain disappeared within 24 hours with 800 mg of parenterally administered cimetidine, and serological data returned to normal.
Titer of anti-varicella-zoster virus antibody rose to 32×.
Cimetidine is known to block the cell mediated immunosuppression by suppressor T-cells with histamine H2-receptors. And, there has been some reports about the treatment of herpes zoster with cimetidine.
Although there have been no reports about the treatment of herpes zoster with cimetidine in Japan, it may become one of most useful agents for treatment of herpes virus infection.

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