Lepromin vs purified bacillus suspension. I. Preparation of a purified bacillus suspension (with a note on nile-blue staining of smears)

The various methods that have been employed for obtaining tissue-free suspensions of leprosy bacilli are reviewed briefly. Most of them were eliminated from consideration for practical purposes, because they deo not meet the primary requirements of simplicity of technique or - and especially - of economy of the leproma material. After experimentation it was found most practical to harvest the bacilli from the pulped leproma tissue with chloroform, as in the first step of preparing Dharmendra's antigen, and then to dilute the pooled chloroform extract with two times its volume of acetone, after which the bacillary bodies are deposited by centrifuging and resuspended in phenol-saline. The bacilli so obtained are affected to some degree by the chloroform, being distinctly paler in color and of thinner appearance after Ziehl-Neelsen staining than those in the standard lepromin made from an aliquot of the same starting material. Consequently, the final suspension was made up with approximately 50 per cent morer bacilli than the lepromin made for use as the control in clinical tests.

Lepromin vs purified bacillus suspension. II. Comparative tests with a purified bacillus suspension

Comparative tests of Wade's "purified acillus suspension" and the standard lepromin prepared from the same pool of leproma have been made in the Philippiones, Netherlands New Guinea, East Africa and South Africa. Consistently in the comparisons the standard lepromin gave more positive Mitsuda reactions, or more strong reactions, than the suspension; and in the healthy children tested the PBS sometimes gave negative results when the reactions to lepromin were positive. For this reason alone it would not be satisfactory for use in field work in which children are involved. Lepromin gave more early reactions than the suspension in the cases for which that effect was reported (Philippines), suggesting that in the preparation of the latter the immediately-available antigenic elements had partly been eliminated. The suspension on the whole caused somewhat less ulceration of the reaction lesions, which is a point in its favor. However, contrary findings were not infrequent, so use of the product could not be advocated strongly on that ground. The results in tuberculoid cases permit comparison of the reactivities of the four different regional groups, whether considering average sizes of the reaction lesions or the percentage of the stronger reactions. Despite the high percentages of positive reactors in the Philippine subjects, the New Guinea people proved to be distinctly more so, and the East African cases the most reactive of all. The South African cases were much less reactive in comparison. The question of why people living in different environments differ materially in reactivity remains an intriguing problem.

A case of advanced borderline leprosy reevaluation of a case originally reported as lepromatous

This article concerns the reevaluation of the classification diagnosis of a Negro patient who had been diagnosed in Pittsburgh, Penna., in 1953 as lepromatous leprosy, which diagnosis had been accepted when he was admitted to the Federal Leprosarium at Carville, La. Actually, the condition was eather advanced borderline leprosy. The primary lesion on the right forearm had at first been thought by the´patient to be an insect bite, but it progressed and when biopsied six months later was reported to be "sarcoid" in nature, which presumably signifies "tuberculoid". When seen at the Aspinwall Veterans Administration Hospital after another eight months, it presented as a mother lesion of the aspect of a major tuberculoid lesion which had undergone borderline deterioration. On the face were many asymmetric small nodular and other lesions which evidently had arisen as an eruption in a tuberculoid reaction, but which had not - and never did - cause loss af the eyebrows. During the month of his first hospitalization there appeared a circinate lesion on the abdomen, described as similar to the one on the forearm. Histologically, the forearm lesion showed a complex picture with a predominance of lepromatous elements, containing an abundance of bacilli in most parts. In the deeper levels of the dermis the picture was definitely lepromatous, but in the main lesion mass in the upper levels the picture indicated that the basis had been a reactiobnal tuberculoid condition, although no distinct tuberculoid features remained. A significant feature of the principal biopsy specimen was the presence, in one end of the section, of a part of an "immune" area against which the active lesion ended abruptly. Such areas arise only by central healing of tuberculoid plaques, and never in lepromas. Although the patient left Carville against advice after only four months, to return to the Veterans Administration for outpatient treatment, his condition has cleared up. This favorable outcome is in accord with the relatively goood prognosis of borderline cases generally.

Results of tests with serial dilutions of lepromin in separate groups of normal young children with a comparison of two lepromins and the Dharmendra antigen

To ascertain whether or not high dilutions of lepromin would be practical for the field testing of normal populations, two series of tests were made with healthy schoolchildren aged 6-9 years. The first series comprised five groups averaging 116 children each, all of Opon poblacion (town), the second one five groups averaging 92 each, all of outlying barrios (rural villages). None had been lepromin tested before or BCG vaccinated, and none was given more than one injection of lepromin. In the first series two lepromins made by different methods (M, by Mabalay, and W, By Wade) were compared with each other and with the Dharmendra antigen, and two dilutions (1/10 and 1/20) of the stock W lepromin were also used. The tests in the second series were with five concentrations of the W lepromin: 1/1 (i.e., undiluted), 1/10, 1/20, 1/40 and 1/80. With respect to the late reaction, the two lepromins in Series 1 gave the same results, 75 per cent positives. The M stock, however, elicited fewer early reactions than the W stock, suggesting that boiling may be less effective than autoclaving in freeing the soluble antigenic elements involved in that reaction. In series 2 the full-dose W lepromin gave rise to more positive reactions of both kinds than it had in Series 1, and the dilutions gave increasingly large differences with respect to the late reaction. The Dharmendra antigen proved ineffective in eliciting either type of response in these very young and immature subjects, this result with the late reaction probably due to the extraction of the lipids during manufacture....