Antigen induced inhibition of cell migration has been used as an in vitro correlate of cell mediated immunity. The capillary method has been used in most studies. More recently, the agarose method was introduced by Clausen, who noted its advantage over the glass capillary technique. The present authors tried to reproduce the Clausen technique for clinical use, but with no satisfactory results. Accordingly, technical variables with this method were reassessed. Following items were drawn from our observation. 1) Bacto agar, instead of agarose, in a concentration of 1% is the most suitable as plate material. 2) The agar medium containing TC-199 and 10% horse serum induces the most satisfactory leukocyte migration. 3) Leukocyte migration area increases with the number of cells in a well. Considering the limited amount of test blood and the defined migration perimeter, the cell concentration of 2×10^6 cells/well seems practically adequate. 4) Addition of horse serum into antigen-leukocyte mixture is not necessary, as the serum added decreases migration area. 5) Opimum timing for maximum migration inhibition comes from 4 to 6 hr after initiation of incubation when stimulated with 50 or 100μg/ml of PPD. More extended incubation time tends to reduce inhibition index. Stimulation with the antigen in a lower concentration slightly enhances migration. 6) Pre-incubation of the antigen-leukocyte mixture before introducing into agar well does not make significant difference in migration inhibition, although prolonged pre-incubation produces a slightly greater migration area. 7) There is no significant difference in the degree of migration inhibition, whenever the same concentration of the antigen is added into the agar plate or into the cell mixture. However, addition of the antigen into the cell mixture is more economical than that into the agar plate.

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In Japan, following kinds of the occupational pollinosis have been reported by some preceding investigators up to now: 1)the sugar beet pollinosis among the research workers of the Sugar Beet Institute, Sapporo (by Matsuyama et al.), 2) the bronchial asthma caused by orchard grass (Dactylis glomerata) pollen among the pasture-cultivators (by Nakazawa et al.), 3) the bronchial asthma induced by strawberry pollen among the strawberry-cultivators (by Kobayashi et al.) and 4) the pollinosis among the pyrethrum-cultivators (by Nakagawa et al.). In this report is described a case of investigator with the occupational pollinosis due to the common ragweed (Ambrosia elatior) pollen, pursuing his study on the same. J.U., a university professor, 56 years of age, is an authority on palynology in Japan, and has been engaged in the research on the common ragweed pollen since 1951. He visited our allergy clinic in April 20, 1970 with the complaint of rhinorroca, sneezing, nasal obstruction, and difficulty of expectoration with slight dyspnoea always after handling common ragweed pollen since 1957. The remarkable peripheral leucocytic eosinophilia was not noticed, but the eosinophilia in nasal smear was observed. In the intracutaneous test with routine inhalatory allergen extracts, a conspicuous positive reaction was proved to the common ragweed pollen and cat-tail pollen, and a middle grade positive reaction to Candida albicans, too. In the nasal provocation test, a positive reaction was obtained with common ragweed pollen and a negative reaction with cat-tail pollen. In September 21, 1974, the Prausnitz-Kustner's passive transfer test with common ragweed pollen extract was carried out, and positive reactions were assured. The serum IgE level at the same time was 5OO unit/ml. Judging from the above-mentioned clinical history and the results of allergic investigations, this case is considered to be the subject with the occupational pollinosis caused by common ragweed pollen. And inferring from this fact, the author would point out the possibility of existence of the occupational allergy among the research workers of the other kinds of pollen, spore or mycelium, and would advocate the need of investigation to them.

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Human blood type O (C^+D^+) red cells sensitized with one of Rh antisera (Ripley) produced firm and stable rosette (Fc-rosette) with some of human lymphoyctes. Fc-rosette was not inhibited by ethylene-diamine-tetraacetate (EDTA) and the effect of incubation temperature was minimal, while aggregated human IgG and Fc fragments markedly inhibited. When lymphocytes are separated into Fc-rosette forming cells and non-rosette forming cells, the former exhibited rosette formation with sheep red cells sensitized with complement and antibody (EAC) and positive stainings with fluorescence labeled antihuman immunoglobulin, whereas the latter produced spontaneous rosette formation with non-sensitized sheep red cells. Reduction and alkylation of Ripley's antiserum abolished Fc-rosette formation. Ripley's antiserum is more effective for the rosette formation than 10 other Rh antisera and rabbit antihuman red cell serum. Positive incidences of Fc-rosette forming lymphocytes from normal healthy adults were 27.1±6.9%, which are similar to those of complement receptor lymphocytes detected with EAC rosette and surface immunoglobulin bearing cells with immunofluorescent method. Under electron microscopic study, Fc-rosette showed distinct features, that is, attachments of lymphocytes to red cells are broader and more intricate than those of other rosettes. These results first indicated that Fc-rosette is produced through Fc-receptors on B-lymphocytes Second, interchain disulfide bonds on IgG molecules are necessary for Fc-rosette formation. Third, human IgG seems to be more effective than rabbit IgG for Fc-rosette formation. In addition, since reduction and alkylation of Ripley's serum abolished Fc-rosette formation and Clq producing cells mainly belong to macrophages, the possibility that either rheumatoid factor producing cells or Clq producing cells are involved in these rosette forming cells is very little. In conclusion, Fc rosette with human red cells sensitized with Ripley's antiserum is a simple and highly specific method for human B-cell identification.

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The types of areal dispersion of air-born pollens were investigated and followed up at four spots located within Niigata City, using by the standard method of pollen counting in the air. Captured pollens in the sampling slides were classified taxonomically and computed in number per square centimeter, and those results were analysed in connection with geographical and botanical differences as well as daily difference during the following up period. Consequently, the type of pollen dispersion was classified into three categories; wide spread (A), intermediate (B) and localized type (C). Pollens in type A were belonged to the anemophilous trees and used to disperse to the places of long distance. Those of type C were belonged mostly to the entomophilous trees and weeds such as reed-mace, dock and ragweeds, and dispersed only within small localized district. While, those of type B were mainly of anemophilous grasses and weeds which dispersed within the range of intermediate distance. Accordingly, the difference of pollen dispersion types should be mentioned for accurate diagnosis, treatment and preventive measures of pollinosis. In addition, geographical distribution of plantation, wind blows and its direction were also important for analysis of influencing factors to patient's residence. In localized type (C) of pollinating, the relation between pollinating and the positive rate of the skin test among asthmatic children was recognized to some extent.

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A case of Chicago acid-induced asthma is reported with several immunological investigations on it. The patient is a 37-year-old man, who has been working in a chemical plant since he was 21-year-old. At the age of 24, he experienced a work to produce Chicago acid, a intermediate product of dye, for 2 months, but during that time he had no abnormal symptoms. Seven years later, he worked again to produce Chicago acid. After half a month of the re-work, he began to complain sneezing, wheezing and paroxysmal dyspnea after exposure to Chicago acid. Although his work changed in the plant, he showed asthmatic symptoms by exposure to another dye, Chicago Blue 6 B. Intracutaneous and Prausnitz-Kustner tests with Chicago acid showed immediate wheal and flare reactions. Provocative inhalation test with Chicago acid produced typical asthmatic attack of immediate type. His complaints disappeared completely by avoiding these antigenic substances. Ten months later, however, his asthmatic attacks recurred by handling S acid, of which chemical structure resembles closely to Chicago acid. These findings suggest that antigenicities of these chemicals may relate to their common chemical structures.

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