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EXPERIMENTAL INOCULATION of HUMANS with ECTODERMOTROPIC VIRUSES* HERBERT GOLDSCHMIDT, Ml)

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EXPERIMENTAL INOCULATION of HUMANS with ECTODERMOTROPIC VIRUSES* HERBERT GOLDSCHMIDT, Ml) View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector EXPERIMENTAL INOCULATION OF HUMANS WITH ECTODERMOTROPIC VIRUSES* HERBERT GOLDSCHMIDT, Ml). AND ALBERT M. KLIGMAN, M.D., Pu.D. Students of human infectious disease have a Vaccinia much greater opportunity to gain an under- To test the hypothesis that freshly regenerated standing of pathogenesis when the disease can beepidermis would promote virus infection, we did experimentally reproduced in man at will. In thisa pilot study with vaccinia in view of the ease of paper we shall describe our efforts to promote thevaccination and the definite knowledge of its inoculation of human skin with the followingcourse. The vaccinations were carried out in the cctodcrmotropic viruses: the wart virus (verruca usual way (multiple puncture technic), using 106 vulgaris and condyloma acuminatum), vaccinia,adult subjects, all of whom had been previously molluscum contagiosum, herpes simplex andvaccinated. The anticipated immune response herpes zoster. A review of previous attempts iswas observed in every instance. In all, 312 inocu- appended. lations were made, one forearm site serving as METHODS the control with the corresponding site on the opposite forearm having been prepared either With the exception of the vaccinia virus, it isby dermabrasion or by cantharidin. The inocula- not at all easy to inoculate humans successfullytions were performed at intervals varying from 1 with the above mentioned viruses. "Takes" areto 18 weeks after preparation. As a standard of infrequent and erratic. Knowing that virusescomparison, the diameters of the lesions (exclu- multiply best in embryonic tissue, the assumptionsive of simple erythema) were measured on the was made that regenerating epidermal cells afterthird day post vaccination. previous removal of the epidermis might in some A condensation of the significant findings is respects be comparable to embryonic cells andgiven in Table I. To show best the enhancing provide a more favorable milieu for virus growth.effect of special preparation, the number and The methods used for this purpose were: a) thepercentage of instances in which the reactions dcrmabrasion technic, and b) the production ofwere at least twice the size of the control sites cantharidin blisters. The dcrmabrasion proce-are recorded. The control sites ranged in diameter dure is the one ordinarily used in removing acnefrom 0.5—1.5 cm. In the prepared sites the range scars in which the skin is first frozen with awas 0.5 to 10.0 cm. The reactions in the derma- refrigerant gas and then planed with a revolvingbrasion sites were consistently larger than in wire brush or serrated stone. Cantharidin blisterscantharidin sites. With the former technique, were raised by introducing 0.25 ml of a 0.5%73% of 87 tests gave reactions at least twice as acetone solution of cantharidin into a glass cupgreat as the control sites whereas the corre- about 1" in diameter. The acetone was evap-sponding figure for the eantharidin site is 36% orated under a stream of air and the site coveredof 88 tests. It will be seen that the size of the with a gauze bandage. Inoculations were madereaction is highly dependent on the time interval into the prepared sites and symmetrical controlafter preparation of the site. With the derma- sites at time intervals ranging from 3 days to 12brasion technic, the optimal time for the maxi- weeks. Healthy colored male volunteers, 20 to 45mum lesion was one month after preparation. years of age, inmates of the Philadelphia CountyAt that time all the inoculations yield lesions at Prison, served as subjects. We mostly used theleast twice the size of the controls. Indeed, the volar surface of the forearm but in recognitionreactions averaged five times the size of the of the great regional variations of the skin, othercontrols. Inoculations performed less than 6 days sites were also tested. after dermabrasion uniformly failed. In sharp *Fromthe Department of Dermatology (Don-contrast to this, the optimal time with the ald M. Pillsbury, M.D., Director), University ofeantharidin technic, was one week, at which time Pennsylvania School of Medicine, Philadelphia,81% of the lesions were at least twice the size of Pennsylvania. Received for publication January 10, 1958. the controls; however, average diameters were 175 176 THEJOURNAL OF INVESTIGATIVE DERMATOLOGY TABLE I passed it through a Chamberland filter. The Vaccinia Inoculations filtrate was inoculated into scarified areas of the upper arm of 3 individuals. After 50 days a Dermabrasion Cantharidin papular lesion developed in one of the inoculated NumberDeemabrasion FCantharidin at sites. A biopsy specimen of this lesion showed weeks atleasttwiceNumber least twice of testsnormal reactionof testsnormalreaction the typical histological structure of a molluseum lesion. 1 15 16 9 (60%) 13 (81%) Wile and Kingery (2) repeated this experiment 2 12 6 (50%) 12 4 (33%) in 1919 with molluseum material passed through 3 19 15 (79%) 18 5 (28%) a Berkefeld filter and injected intraeutaneously. 4 11 11 (100%) 11 2 (18%) 5 12 8 (66%) 12 2 (20%) Papules developed in the inoculation areas after 6 7 6 (86%) 7 1 (14%) 2—3 weeks and typical molluseum lesions were 7 — — — — present by 8 weeks. 8 2 2 (100%) 2 2 (100%) Findlay (3) repeated these experiments using 9 4 3 (75%) 5 2 (40%) molluseum material stored in 50% glycerol for 10 — — — — 1 month. 11 — — — Neither Pinkus (4) nor Blank and Rake (5) 12 2 2 2 1 were able to induce experimental molluseum 13 1 1 1 0 lesions. 14 — — — — 15 — — — — Present study 16 — — — — 17 — — — — Using 3 different sources (children under 10 18 2 1 2 0 years of age) cores of molluseum lesions were ground in 1 cc of normal saline and inoculated Average: 87 64 (73%) 88 32 (36%) into scarified normal forearm skin and into sites prepared by dermabrasion and eantharidin. usually 1—2 cm less than in dermabrasion sites.In addition, a number of inoculations were made The dermabrasion technic clearly gave superiorinto the axillae. A total of 141 inoculations (60 results by these standards. In accounting forof them at prepared areas) were performed in 30 these differences, it should be recognized thatindividuals. In no ease did a typical molluseum dermabrasion is a far more damaging procedurelesion develop during an average observation which completely removes the epidermis, whereasperiod of 6 months. a layer of epidermal cells subtends the bullae produced by eantharidin and the inflammationComment is less intense. Perhaps an optimal period of 4 Although we had no success, we were able to weeks with the dermahrasion technic as comparedconfirm the observation of Pinkus (4) who de- to one week with eantharidin indicates approxi-scribed a papular "immune" (tuberculin type) mately similar stages of regeneration at thesereaction after inoculation of molluseum material. times. Blank and Rake (5) noticed similar reactions. Subsequent studies with other eetodermo-In 21 of 30 individuals we observed erythema, tropic viruses utilized these optimal times. edema and marked induration developing in one or several of the inoculated areas, the maximum Melluscum contayiosum reaction occurring between the 3rd and 5th day The infectious nature of molluseum eontagio-and disappearing after 7 to 10 days. Almost all sum has been suggested by many reports of acci-of these reactions occurred in normal skin areas dental transmission of the disease by direct hu-and only very rarely in areas prepared by derma- man contact. Attempts to induce molluseum le-brasion or eantharidin. The explanation given by sions in laboratory animals have been unsuccessful. Pinkus and Blank and Rake for these delayed Juliusberg (1) was the first (1905) to establishtuberculin type responses seems very attractive. the viral nature of the causative agent experi-They occur in persons rendered hypersensitive mentally. He ground the cores of molluseumby a previous inapparent infection. If this as- lesions, suspended the material in broth andsumption is correct, the great majority of adult EXPERIMENTAL INOCULATION OF HUMANS 177 persons have sustained inapparent primary infec-5 individuals with material from a papillomatous tions; only a small fraction of the populationwart of the scalp were unsuccessful. ever develop clinically apparent molluscum le- In 1924 Serra repeated his experiments using a sions. Furthermore, it may well be that inappar-filtrate of papillomatous warts of the scalp and cnt infections which render the host allergic maythe suprapubie area. Six of 18 inoculations in 3 also render it immune. From the kind of data wevolunteers were positive, all of them occurring on have given, it is impossible to tell whether thethe dorsum of the hand. Four of these positive failure to produce manifest molluscum lesions isinoculations were observed in the subject from due to a preexisting immunity or whether anwhom the original material was taken. Inocula- actual infection occurred which was inapparcnt.tions of the skin of the penis and the scalp were One way to attack this problem is to find out ifnegative. previously non-allergic subjects become hyper- Tueeio (10) induced warts in the second genera- sensitive after inoculation with live virustion. He used ground material from verrueae Finally, it should be appreciated that otherfiliformes of the beard area and inoculated totally unknown forces may determine whetherscarified areas of the upper arm of 5 individuals infection, apparent or inapparent, will occur.under 18 years of age and of his own arm. Warts Unfortunately, we did not determine whether thedeveloped in 4 of these areas after 5—6 months. "tuberculin" type response was purely allergicOne of them was used to produce a second genera- by using killed virus. If live virus is required, thetion wart. reaction is truly one of immunity similar to the Wile and Kingery (11) used material from immune response in ordinary vaccination. several warts, passed it through a Berkefeld filter and injected the filtrate intraeutaneously Verruca vulgaris into the dorsa of hands of 3 individuals.
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