Comparative Experimental Study of Sensitivity to Tuberculin and
Transcrição
Comparative Experimental Study of Sensitivity to Tuberculin and
Vo lum e .18. ' 1' ''1 t: R:,\A 1I ON,\L JO URNAL 01' 1. 1·: I'RO.'1 ' · l~rilltrd ~' lImh cr 4 i" tlte U .S.A . Comparative Experimental Study of Sensitivity to Tuberculin and Lepromin in Guinea-pigs Previously Inoculated with BCG by Oral and Intradermal Routes 1_ R. D. Azulay and "de Kahn 3 Premunition by BCG for prophylaxis of leprosy and tuberculosi s has been tri ed by several routes of inoculation. Regarding th e intradermal route, Brown et oZ. (] 1 ) showed that BCG protects against leprosy, and D'Arey Hart et a7. ( 12) showed protection against tuberculosis. In Brazil, oral BCG premunition has b:'c n ca rri ed out in several million children sin ce 1927. The value of the oral route with respect to positive conversion of the tuberculin and lepromin tests has been presented in several publications either in th e field of leprosy ("I. 16 , 18) or in th e fi eld of tuberculosis (1.R.15,17). There has been discussion concernin g th e immunologic and practical advantages of one route over others (9, 10). This problem is very important with respec t to costs of administration and determination of th e most practical u se of BCG premunition in leprosy and tuberculosis in mass campaigns in und erdeveloped countries. A positive lepromin t est is regarded as a sign of resistan ce against leprosy and a positive tuberculin test is an indirect sign indicating th e presence of some resistance against tu bercu losis despite recognition of the fact that tuberculin allergy and resistance against tuberculosis are different phenom ena. Ncverth elcss, post-vaccin ation all ergy is the onl y sign which p ermits evaluation , in children. of the effi ciency of premunition. In elucidation of the problem of th e optimum route of inoculation , an investigation of th e lepromin and tubercu1 R ece ived for pllbli ca ti on 2'1 Ma rch 1969 . ~ Paper presented 10 Ih e I Xl h Internali on al Le p. rosy Congress, Londo n , 16·2 1 Sepl embel' 1968. 3 R . D. AZlda y, M.D ., Chi ef of In st itllte of Lep· rology, Nat ion al Service of Leprosy, Brazil , and I'l'Ofe~sor of D e rm a l.o l o~ y a t U ni ve rsid ade Fed eral F1l1minell se a nd at U ni versid ade do Estado da G u a nab ara; H . Kahn , M .D .. Direc tor of Fllnd a((ao Al:l llifo de P a iva. nCG Service. Brazil. lin tests after oral and intraderm al inoculation of BCG in guin ea-pigs was undertaken. ( A similar study is in progress in new born infants and children. The results will be presented later.) This paper shows th e 6rst results of the experim ent; th e 6nal results will be published after th e verifi cation of lepromin and tuberculin sensitization as evidenced by th e tests don e fi ve months after BCG .inoculations. MATERIALS AND METHODS Materials. One hundred guin ea-p igs, thrce to four months old, wcighin g approximately 350 gm. each, were used. Vaccinations were made with either lyophilized BCG produced by Glaxo Laboratories, Greenford, England ( Batch B818 J) , or BCG Moreau ( Rio de Janeiro strain ), maintained altern atively in glycerinepotato and bile-potato media . . Tuberculin hypersensiti vity was tes ted with Old Tuberculin ( OT ) and PPD by intradermal route. Lepromin hyp ersensiti vity was t ested with lepromin antigen, Hayashi-Mitsuda type ( 16 X 107 bacilli per ce. ). Technic of vaccination. All the 100 guinea-pigs were negative to an initial tuberculin t est. They were di vided in four groups (A, B, C and D ), each havin g 25 guinea-pigs. Group A-Control group. Group B-Intraderm al inoculation with 0.05 mg. of lyophilized BCG. Group C- Intradermal inocul ation with 0.05 mg. of BCG Moreau ( Rio de J anei1'0 strain ) . Group D - Oral inocul ation with 100 mg. of BCG Moreau ( Rio de Janeiro strain ), The inocula used were determined by analogy with those used in newborn infants 373 1970 International Journal of Leprosy 374 '!. 100 90 L[ P "' ". z <r 0- 10 0 8 REACTIVITY F.I G. POSITIVITY 1. Results of tes ts to tuberculin, PPD, and lepromin after vaccination with BCG . Reacli vity : reactions having 1 or more mm. in diam eter. Positi vit y: reactions havin g 2 Dr more mm. in di ameter. as recomm end ed by Glaxo Laboratories and Viscondessa de Morais's Institute, Rio de Jan eiro. Azu]ay et ol. (5-8) showed that BCG oral vaccination in doses of 30 mg. per kg. of animal weight induces 87 p er cent lepromin positivity in guinea-pigs. Amaral and Sorensen (2) obtained 90 per cent positive tuberculin test and in newborn guinea-pigs after administration of 20 mg. BCG by mou th ( 13, H). Thirty days after vaccination all guineapigs were t ested with OT, diluted to 1/ 10 units of PPD ( Ht 23-Copenhagen) and TA BLE 1. Results uf tests to tuberculin, P P D and lepromin after vaccination (cith BGG. R eactivity" ....'" P PD B(,G ('ontrol BCG % % % % 22 .7 78 .7 39 .3 22 61 GLA)/:O INTRAOE.ANIC - G,.OUP C l eG UP NOR[AU ' INTRAO[Mt IC w - - - TU' ...... - - .rpo _ GROUP 0 L[I' leG WOMau - OAAL 73.8 16 . 3 Lc~m:' ____ I--=~=- ~~~ _~ 5 ~~~ No. guin ca-pig,.; BeG - Positi vityb tl'ol Tuberculin , --- ~ - . - TU' -----------('on- GROUP 8 ~ 22 61 I " H,eadivit y = reactions having or more n1l11. in diameter. b Positivi ty = rea ctions hav ing 2 or more mm. in diameter. I ..~---+ -- .. -.. _- : O "~ Z 3 . - . -. "1'0 _ . LEI' FIC. 2. Sensitivity ,to tuberculin, PPD , and lepromin in guinea-pigs previously inoculated with BeG. 1 I 1 1 1 59 .1 27 .2 9.1 1 13 .6 27 .2 27 .2 1 1 59 .1 27.2 100.0 1 14 .2 100 .0 85 .0 25 .0 85 .7 1 76.1 28 .5 90.4 1 33 .3 85 .7 Group D. BCG Moreau oral 59 . 1 27 .2 81.8 I 1 Group C. BCG Moreau intradermal 63 .6 13 .6 4 .5 Group B. BCG Glaxo intradermal 1 22 .7 27.2 - 1 9.5 90.4 I. 76.1 - 50 .0 - 70 .2 9.1 - - - 1 1 1 1 21 Tuberculin PPD Lepromin Tuberculin PPD Lepromin Tuberculin PPD Lepromin Tuberculin PPD Lepromin Control 1 I I II 1 I 7 - 1 4 .5 - Grou p A . Control 72 1 1 1 54.5 27.2 4 .5 1 I 54 .1 9.1 100 .0 1 80 .0 10 .0 85.0 1 76.1 14 .3 90.4 I 1 100 .0 - 14 .2 80.7 - 33 .3 I I 7G. 1 90.4 - 50 .0 - 4 .5 ..... ~ en '" ~. -d >=l (\) S· ~ ;;. ;;. ;3 d ""13 (\) t"'< G- S- .: ~ (\) 0'" ~ o ;i.: <>::: g.~ (\) en ;:;-;::s ~ G- <>::: ~ ~ ~ >I>- - - w yO? - - 1 1 1 1 1 21 - 9 .5 70 .2 - - - 14 Days intrader~zal 13 .6 9 .1 27.2 Group D . BCG .1foreau oral 59 . 1 4.5 63 .5 Grou p C. BCG J[oreau 54 .5 13 .6 4 .5 Group B. BCG Glaxo intradermal - 48 Hours % Positivityb PPD, and tuberculin, after BeG vaccination by different routes. - - - 14 Days to lepromin, • Reactivity = reaction having 1 or m ore mm. in diameter. b Positivi ty = reaction having 2 or more mm. in diameter. Tuberculin PPD Lepromin Tuberculin PPD Lepromin Tuberculin PPD Lepromin - - - - - I 7 Group A . Control Tuberculin PPD Lepromin 1 72 48 Hours % Reactivitya 2. Sensitivity of guinea-pigs Control TABLE 376 I ntemational }ollmal of Leprosy lepromin containing 16 X 107 bacilli/ ml. by intradcrmal route. The readings "vere mad e at 48 hours, a,ncJ at 7, 14 and 21 days. Criteria for reading results. Positivity was determined by the frequency of the largest reactions measured in millimeters. A "reactive" test means any reaction independent of th e size of it. The criteria of positivity were as follows: 1. Positive tllJJercuIin test : reaction of 2 or more mm. at 48 hours. 2. ·Positive PPD test: reaction of 2 or more mm . at 72 hours. 3. Positive lepromin test: reaction of 2 or more mm. at seven days. RESULTS Group A (control ) 1970 Group D (oral BCG.Morcau). 1. Tuberculin test: 80%positive; thi s decreased to 76.1%, .33.3% and 9.5% at the 72 hour, 7 day, and 14 day readings respectively. Absence of any reaction at 21st day reading. 2. PPD test: 14.3%positive but th e I'eae" tivity was twi ce as hi gh (28.5%). Absencc of any reaction at 7, 14 and 21 days, respcctively. 3. Lepromin test: 85.0% positive; this increased to 90.4% at th e 14th day reading and dccreased to 76.1% at the 21st day readin g. SUMMARY AND CONCLUSIONS A definite sensitization of guinea·pigs to Old Tuberculin , PPD and lepromin was 1. Tuberc ulin test: absence of any reacinduced by the adminis·tration of BCG. The tion 2. PPD tes t: no positive test, but some reactivity reactions were a little bit higher discrete reactivity ( 1 mm. ) was than the positivity reaction s. The hi ghest points of the curves of posipresent in 22.7%at seven days. 3. Lepromin test: 4.5% positive test and tivity were: 48· hours for th e tuberculin test, 72 hours for th e PPD test and seven 27.2%reactive tes t at seven days. days for th e lepromin test. Group B (intradermal BCG.Glaxo). The lepromin reaotion in guinea·pigs is of shorter duration than in human beings. 1. Tuberculin test: 54.5% positive and The induced sensitization of th e guinea63.6% reactive test. It is interestin g to observe that 13.6%of the positive tests pigs varied according to the type and route were still evident at th e seventh day. of administration of BCG. At the later readings all reaction s had High est tuberculin sensitization was disappeared. achieved in Group D (oral BCG-Moreau ). 2. P<PD test: 27.2% positive; 9.1% rePPD sensitization was high est in Group mained positive until the seventh day. B (intradermal BCG-Glaxo). 3. Lepromin test: 27.2% positive; 4.5% Lepromin sensitization was very low in remained positive until the 21st day. Group B ( intradermal BCG-Glaxo ); it was very high in Group D ( Oral BCG Moreau ) Group C (intradermal BCG·Moreau). and th e hi ghest in Croup C ( intradermal 1. Tuberculin test : 59.1% positive; 14.2% BCG-Moreau) . remained positive until the seventh In Groups C and D , which received day; at later readings all reaction had BCG-Moreau, the lepromin reactions were disappeared. more p ersistent; in the Group D ( oral BCG 2. PPD test: 9.1% positive, but the reac- Moreau ) the reactions were even more tivity was three tim es higher (27.2%); persistent than in th e Group C ( intraderabsence of any reaction at later read- mal BCG-Moreau). ings. As th e oral administration of BCG is 3. Lepromin test: 100%positive; this de- easier and cheaper it should be recomcreased to 70.2% and to 50% at the mended in the under developed countries, 14th and 21st day readings, respec- in the prophylaxis of leprosy and tubercutively. losis. 38,4 Azulay & Kahn: Sensitivity to Tuberculin & Lepromin in Guinea-pigs RESUMEN La administracion de BCG indujo en cobayos un a sensibilizacion bi en definida a tube rculina antigu a, PDD y lepromin a. Las reacciones de re ac tividad fucron ligeramente mayo res que las reacciones de positividad . Los puntos mas altos de las curvas de positivid ad fueron : 48 horas para la prueba de tuberculin a, 72 ho ras para la prueba con PPD y siete dias pa ra la pru eba con 1epromina. La reaccion de lepromin a en co bayos es de menor duracion que en seres hum anos. La sensibilizac ion que se indujo en los cobayos vario de ac uerdo co n los tipos de BCG utilizados y vias de administracion . La mayo r sensibili zacion contra la tuberculin a se obtuva en el Grupo D (BCG oralMoreau). La sensibilizacion contra PPD fue mayor en el Grupo B (BCG intradermico- Glaxo) . La sensibilizacion contra la lepro min a fue muy baja en el Grupo B (BCG intradermicoGlaxo ); mu y alta en el Grupo D ( BCG oral - Moreau) y mayo r de tod as en el Grupo C (BCG intrade rmico- Moreau) . En los Grupos C y D , que recibieron BCG -Moreau, las reaccion es de lepromin a fueron mas persistentes; en el Grupo D (BCG oralMoreau) las reacciones fueron allll mas persistentes que en el Grupo C (BCG intradermico - Moreau). Ya que la administracion oral de BCG es mas facil y de menor costo, debe se r recomendada para paises en desa rrollo, para la profilaxis de la lepra y la tuberculosis. 377 La sensibilisation par Ie PPD s'est revelee la plus elevee dans Ie groupe B (BCG intradermique Gl axo). La sensibilisation par la lepromine etait tres fa ible dans Ie groupe B (BCG intradermique Glaxo); elle etait tres elevee d ans Ie groupe D (BCG Oral Moreau ) et la plus eleves dans Ie gro upe C (BCG intradermique Moreau) . Les reaGtions a la le promine sont apparues les plus persistantes dans les groupes C et D qui ava ient recru du BCG Moreau ; pour Ie groupe D (BCG oral Morea u ); les reaction s eta ient meme encore plus persistantes que dans Ie groupe C. (BCG intradermique Moreau ) . L'administration de BCG ora l dev rait etre recomm andee pour la prophylaxie de la lepre et de la tuberculose dans les pays en voie de developpement, vu qu'elle est plu s facile et plus economique. REFERENCES 1. ALvIl"r, A. C. and NASCIJ\IENTO, E. A. D ez anos de vaci nac;:ao BCG concorrente na Favela d a Praia d o Pinto. J. Brasileira Doene. T onie. 2 ( 1966 ) 8-9. 2. A",rARAL, J. P. an d SOERENSEN, B. BCG pol' via oral em eob aios. 0 H ospital 58 ( 1960 ) 1053-1057. 3. ASSIS, A. PASSADO, presente e futuro da vaeinac;:ao BCG. Rev. Servo Nae. Tubere. 8 (1964) 7-30. 4. AZULAY, R. D . A aean do BCG sobre a reac;:ao le promini ea. 0 H ospital , 34 ( 1948) 853-856. 5. AZULAY, R. D . Our exp eri ence w ith the leprom in ·t est from 1944 to 1955. II Congo RESUME Intern . Alergia, Rio de Janerio, 1955. L'administration de BCG a entraine une 6. AZULAY, R. D. and AZ ULAY, J. D. A virasensibilisation indi scutable de cobayes a la gem da lepromin o-readio pela adminisVieille Tuberculine, au PPD et a la lepromine . trac;:ao do BCG. 1301. Servo Nae. Lepra 12 Les reactions de r eac tivation etaient (1953) 310-312. legerement plus prononcees que les reactions 7. AZ ULAY, R. D. and NEVES, R. G. Comde positivite. p ortamento do teste leprominieo em Les points les plus eleves des courbes de eobaios beeegeizados p Ol' via oral. Rev. positivite obtenues etaient les suiv ants; 48 brasileira L epro!. 2 6 ( 1958 ) 103-106. heures pour I'epreuve a la tuberculine, 72 8. AZULAY, R. D ., NEVES, R. G. and Az ULAY heures pour I'epreuve au PPD et 7 jours pour .T . D . A reac;:ao le prominica apos previ~ I'epreuve a la lepromine . in oculac;:ao do BCG e de Mycobacterium La reaction a la lepromine chez les cobayes tllberculosis morto p OI' irradia<;ao. Rev. temoignait d' une duree plus cOllrte que chez les . brasileira Lepro!. 27 ( 1959) 81-86. etres hllmains . 9. BLASI, A. and CuRCI, G. Investigac;:oes exLa sensibilisation ainsi indllite chez les p erimentais sabre a vacinac;:ao com BCG cobayes variait en fonction du type de BCG pOI' vi a oral segundo A. de Assis. 0 Hoset de son mode d'administration. pita l 47 ( 1955) 215-229. La sensib ilisation la plus forte a la tubercu10. BOTES, V., PARASCHJVESCO, M ., ANGELESline a ete obtenue dans Ie groupe D (BCG CO, 1. , BONCIU, C. and PETROVICI, M. Moreau Oral) . International l ou'm al of L eprosy 378 E studo exp e rimental sabre a vaci na9ao BCG intensiva pro via oral. 0 H ospital 61 (1962 ) 441-45Q. 11. BROWN, J. A. 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Considera90es sobre 0 BCG liofili zado. 0 H ospital 67 (1965) 215- 237. 16. ROSE}'fBERC, J., SOUZA CAMPOS, N. and A UN, J . N. Rea9ao de Mitsuda induzida p OI' efcito d e di ve rsos esquemas d e vacina9ao BCG oral e p ela tecnica de multipuntures de Rosenth al. Rev. brasileira L ep ro l. 20 (1952 ) 183-196. 17. SrCNORINI, L. F . Esperie nze di vaccinazione antituhercol are dell a vacia pe r via orale. Lo Sperimentale 106 (1 ) (1956 ) 88-100, and 0 H ospital 50 (2 ) ( 1956 ) 161-162. 18. SOUZA CAMPOS, N ., L ESER, 'vV ., QUAGLIATO, R. , BECHELLI, L. M., and ROTBERG, A. Viragem da lepromino-rea9ao em fun 9ao de diferentes es timulos. Influencia da idad e, nessa v irage m, no grupo etario de 6 a 43 meses. Rev. brasil eira L eprol . 29 (1962 ) 3-20.