Assessment of immunological markers and booster effects of Ag85B peptides, Ag85B, and BCG in blood of BCG vaccinated children: a preliminary report

Aliabbas A. Husain,Hatim F. Daginawla,Lokendra Singh,Rajpal S Kashyap 대한백신학회 2016 Clinical and Experimental Vaccine Research Vol.5 No.1

Purpose: In the present study, the protective immunological markers in serum and peripheral blood mononuclear cells (PBMCs) of bacillus Calmette–Guérin (BCG) vaccinated and unvaccinated children were evaluated after vaccination. Further, PBMCs of children with low protective levels were boosted with BCG, Ag85B, and Ag85B peptides to study their booster effects to increase waning BCG induced immunity. Materials and Methods: Fifty children from 1 month to 18 years of age were randomized for the study. Blood samples were collected from 27 participants with/without BCG vaccination. Immunological markers (anti-BCG, interferon γ [IFN-γ], and adenosine deaminase activity) were assessed in both serum and PBMCs of children. Children with low levels of protective immunological markers were further recruited and their PBMCs were boosted with BCG, Ag85B, and Ag85B peptides. Results: Children in age group of 4-6 years were associated with significantly (p<0.05) higher BCG-specific IgG and IFN-γ levels compared to those in age group greater than 10 years. Vaccinated children had greater repertoire of immunological memory which on in vitro stimulation with BCG showed increase in BCG-specific response compared to unvaccinated controls. Assessment of booster effects of BCG, Ag85B, and Ag85B peptides in PBMCs of children revealed greater potential of peptides to boost BCG induced immunity compared to BCG and Ag85B. Conclusion: To conclude, children within age 4-6 years are associated with high immunological markers which eventually diminish with age thereby suggesting need for booster dose in later years. Mycobacterium tuberculosis peptides along with BCG may be used as attractive candidates to boost such waning BCG induced immunity in children.

BCG가 Ehrlich 암세포를 이식한 생쥐의 위점막 상피세포의 DNA합성 및 미세구조에 미치는 영향

고정식,류인상,박경호,박대균,Ko, Jeong-Sik,Ryoo, In-Sang,Park, Kyung-Ho,Park, Dae-Kyoon 한국현미경학회 2009 Applied microscopy Vol.39 No.3

이 실험은 Ehrlich 종양세포를 이식한 후 BCG를 투여하였을 때, 위점막 상피세포의 형태학적 변화와 DNA합성능의 변화를 연구하고자 시행하였다. 실험동물로는 체중 25 g 내외의 성숙한 생쥐(ICR계통)를 정상대조군, 종양세포이식대조군(종양대조군), 종양세포이식후 BCG투여군(BCG투여군)으로 구분하였다. 종양대조군과 BCG투여군 동물들은 샅부위 피하에 각각 $1{\times}10^7$의 Ehrlich 종양세포를 이식한 후, 다음날부터 BCG ($0.6{\times}10^8{\sim}6.4{\times}10^8$ CFU, 27 mg/vial, Connaught Lab., Canada)를 하루건너 한 번씩 피부밑조직에 주사하였으며, 종양대조군은 종양세포이식 후에 BCG 대신 0.2mL의 생리식염수를 피부밑조직에 주사하였다. 자기방사법적 관찰을 위해서는 BCG를 마지막으로 주사한 다음날 $^3H$-thymidine (methyl-$^3H$-thymidine: specific activity 25 Ci/mmol, Amersham Lab., England) 0.7${\mu}Ci/gm$를 꼬리정맥에 주사하고, 70분 후 도살하여 위조직을 떼어내어 10% formalin에 고정하였다. 자기방사표본관찰은 위점막 조직이 세로로 잘 절단된 부위를 택하여 점막근육판을 따라 점막길이 3.5mm의 위점막 조직에 분포하는 $^3H$-thymidine 표지세포의 수를 계수하였으며, 일반조직 관찰을 위해서는 hematoxylin-eosin (H-E)염색을 시행하였다. 전자현미경 관찰을 위해서는 떼어낸 위조직을 2.5% glutaraldehyde-1.5% paraformaldehyde 혼합액에 고정한 후, 1% osmium tetroxide 액에 다시 고정하였으며, 고정이 끝난 조직은 탈수과정을 거쳐 araldite 혼합액에 포매하였다. 광학현미경적 관찰에서 종양대조군과 BCG투여군은 위점막 조직에서 형태적으로 큰 변화를 볼 수 없었다. 전자현미경적 관찰에서 BCG투여군의 점액상피세포는 전체적인 모습이 정상대조군과 종양대조군의 소견과 유사하였으나 정상대조군과 종양대조군에 비해 수초구조와 뭇소포체(multivesicular body) 및 전자밀도가 높은 큰 미토콘드리아속과립이 자주 관찰되었다. 자기방사법적 관찰에서 정상대조군, 종양대조군, BCG투여군은 점막길이 3.5 mm 당 출현하는 표지세포수가 각각 380.2 (${\pm}31.35$), 426.1 (${\pm}28.43$) 및 301.8 (${\pm}34.63$)개이었으며, BCG투여군은 정상대조군과 종양대조군에 비하여 표지된 은입자의 수가 매우 적어서 표지과립이 겨우 구별될 정도의 세포가 많이 관찰되었다. 이상의 결과를 종합해보면 Ehrlich 종양세포를 이식한 동물에 BCG를 반복 투여하면 위점막 상피세포의 DNA합성을 효과적으로 억제하면서도 형태적인 변화가 매우 경미하였으며, 이러한 결과는 BCG가 항암치료 시 보조제로 사용할 수 있는 좋은 약제라고 생각된다. This experiment was performed to evaluate the morphological responses of the gastric epithelial cells of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of BCG. Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups (tumor control group and BCG-treated group). In the experimental groups, each mouse was inoculated with $1{\times}10^7$ Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day after inoculations, 0.2 mL of saline or BCG (0.5 mL/25 g B.W.: $0.03{\times}10^8{\sim}0.32{\times}10^8$ CFU) were injected subcutaneously to the animals every other day, respectively. The day following the 7th injection of saline or BCG, each mouse was injected with a single dose of 0.7 ${\mu}Ci/g$ of methyl-$^3H$-thymidine (25 Ci/mmol, Amersham Lab., England) through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed, and gastric tissues were taken and fixed in 10% neutral formalin. Deparaffinized sections were coated with autoradiographic emulsion EM-1 (Amersham Lab., England) in a dark room. The number of labeled epithelial cells in the gastric mucosae (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and calculated. And for electron microscopic observation, gastric tissues were prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by post-fixation with 1% osmium tetroxide solution. On the light microscopic study, gastric mucosae had no morphological changes following the injection of BCG. On the electron microscopic study, in the BCG-treated mice, myelin figures and multivesicular bodies within the gastric epithelial cells were observed more frequently than in those of the normal control ones. On the autoradiographic study, number of the labeled cells of normal control, tumor control and BCG-treated mice were 380.2 (${\pm}31.35$), 426.1 (${\pm}28.43$) and 301.8 (${\pm}34.63$), respectively. In the BCG-treated mice, poorly-labeled cells containing only a few silver grains of 3H-thymidine were observed more frequently as compared in those of the normal control and tumor control ones. From the above results, BCG may suppress the DNA synthesis of the gastric epithelial cells, but does not results severe fine structural defect on the gastric epithelial cells. These results suggest that BCG is expected as one of the effective supplemental anticancer drugs.

N-butyl-N-(4-hydroxybutyl) Nitrosamine에 의해 유도된 흰쥐 방광암에서 L-arginine 복용이 방광 내 BCG주입요법의 항암 효과에 미치는 영향

조명근,김종성,임정식 大韓泌尿器科學會 1999 Korean Journal of Urology Vol.40 No.5

BCG 접종 후 국소 림프절염의 임상양상

배선영,박양준,김종현,오진희,고대균,강진한,Bae, Sun Young,Park, Yang Joon,Kim, Jong-Hyun,Oh, Jin Hee,Koh, Dae Kyun,Kang, Jin Han 대한소아감염학회 2006 Pediatric Infection and Vaccine Vol.13 No.2

목 적 : BCG 접종 후 가장 흔하게 발생하는 이상 반응인 국소 림프절염이 동반된 소아들을 대상으로 이들이 접종받은 BCG 균주의 비율, 림프절 크기에 따른 화농화 빈도, 약물치료 여부에 따른 임상 경과를 조사하여 이에 대한 발생 요인을 찾고, 치료방향을 설정하는데 도움이 되고자 본 연구를 시도하였다. 방 법 : 1997년부터 6년의 기간동안 가톨릭대학교 성빈센트병원 소아과 외래에 내원한 환아 중 BCG 접종 후에 발생한 국소 림프절염으로 진단된 생후 24개월 미만의 영유아 52명을 대상으로 하였다. BCG 균주의 종류, 접종 장소, 림프절염의 위치와 크기를 의무기록을 통하여 후향적으로 확인하였고, BCG 균주, 림프절염의 크기, 자연 치유율 및 화농화간의 상관관계를 조사하였다. 결 과 : BCG 접종 후 국소 림프절염의 첫 발견시기의 연령은 평균 5.5개월이었으며, 내원시의 림프절 크기가 클수록 첫 발견시기의 연령이 어렸고, 발생부위는 BCG를 접종한 같은 쪽의 겨드랑 부위가 가장 흔하였다. 대상군 52례 중 Pasteur 균주를 피내 접종한 군이 46례(88.5%)로 대부분을 차지하였고, Tokyo 균주를 경피 다천자법으로 접종한 군은 5례(9.6%)이었다. 이 중 28례(53.8%)는 자연 치유되었고 24례(46.2%)는 화농되었는데 림프절의 크기가 클수록 화농 빈도는 의미있게 높았으나, 균주에 따른 차이는 없었다. 약물치료는 화농 예방에 효과가 없었으며 치유기간을 단축시키지도 못했다. 결 론 : BCG 후의 림프절염의 발생은 균주와 접종방법에 따른 차이가 있는 것으로 예측된다. 약물치료는 효과가 없었으므로 권장되지 않으며 특히 비화농성인 경우는 자연히 소실될 수도 있기에 치료없이 관찰만이 요구된다. 이상적인 BCG는 접종 후 반흔을 남기고, 결핵 피부반응검사에 영향을 덜미치면서 이상반응이 적게 나타나며 저렴한 것이다. 이러한 조건을 가장 많이 충족시키는 BCG 균주를 선택하여 사용하는 노력이 필요하다 하겠다. Purpose : A regional lymphadenitis is the most frequent adverse reaction of BCG. In order to find out developmental factors and establish a strategy of management, we investigated the clinical courses of children with lymphadenitis following BCG on the aspect of BCG strains, suppurative rates according to the sizes of lymph node and the clinical difference with or without treatment. Methods : From January 1997 to June 2004, 52 children less than 24 month-age-old diagnosed as BCG lymphadenitis in Department of Pediatrics, St. Vincent's Hospital, The Catholic University of Korea were enrolled. The type of BCG strain, place of vaccination, location and size of lymphadenitis were assessed with medical records, retrospectively. Finally, we analysed the correlations between BCG strains or the sizes of lymph node and natural remission or suppuration. Results : The first detected mean age of BCG lymphadenitis was 5.5 month-age. The larger of the measurement was at the first visiting, the younger of age that was first presented. The most frequent location was the same sided axillary region of BCG injection. Among 52 subjects, 46 cases(88.5%) were vaccinated with intradermal Pastuer strain, and only 5 cases(9.6%) were done with percutaneous multipunctured Tokyo strain. Twenty eight cases(53.8%) were regressed naturally, otherwise 24 cases(46.2%) were suppurated. The larger those were sized, the higher freqeuncies those were suppurated on, significantly. Treatment with medications could not prevent the suppuration and could not shorten the healing periods. Conclusion : We predict that there are differences between the occurrent rate of BCG lymphadenitis and BCG strains or methods. Treatment with medication is not recommended owing to its ineffectiveness. Especially, in case of non-suppurative lymphadenitis should be onlyless influence on the tuberculin skin test, cause less adverse reactions, and is inexpensive. observed without treatment, because it could be regressed naturally. An ideal BCG makes a scar, We should make an effort to choose the best BCG strain that can fulfill such requirements.

유아의 원위 요골부에 발생한 Bacille Calmette-Guérin (BCG) 골수염

김지영,전지인,장학 대한수부외과학회 2020 대한수부외과학회지 Vol.25 No.2

BCG (Bacille Calmette-Guérin) vaccine has been administered safely to billions of people all over the world. The Tokyo-172 strain has reported to have a lower virulence and side effects than other strains. BCG osteomyelitis of distal radius is a very rare but serious complication due to generalized dissemination of BCG. We report a rare case of BCG osteomyelitis of the distal radius in a 21-month-old girl who had no underlying disorders. Although uncommon, BCG osteomyelitis should be considered a possible complication of BCG vaccination under certain clinical features for early diagnosis and proper treatment. BCG 백신은 전세계적으로 널리 접종되고 있는 백신 중 하나로 그 중 Tokyo-172의 경우 비교적 안정하고 부작용이 드문 것으로 알려져 왔다. 원위 요골의 BCG골수염은 BCG 접종의 드문 합병증으로 BCG 균의 전신성 파종에 의해서 생긴다. BCG 접종에 의한 골수염은 드물지만 심각한 합병증을 유발할 수 있으며, 빠른 진단과 적절한 치료를 위해 감별진단으로 고려되어야 한다. 저자들은 생후 1개월에 BCG 접종을 받은 21개월된 여아에서 좌측 원위 요골부에 발생한 결핵성 골수염을 진단하였기에 보고하는 바이다.

BCG 접종후 Mantoux 검사의 반응도와 임상인자들의 영향

이준성(Joon Sung Lee),이현정(Hyun Jung Lee),김영훈(Young Hoon Kim),황경태(Kyung Tai Whang),김종완 대한소아알레르기호흡기학회(구 대한소아알레르기 및 호흡기학회) 1997 소아알레르기 및 호흡기학회지 Vol.7 No.2

목 적 : 소아에서 Mantoux 검사는 결핵 감염의 유무를 판단하는 데 널리 사용되는 방법으로, BCG 접종후 면역 상태를 알아보는 데도 유용하다. 그러나 그 반웅의 정도 및 양전율은 백신접종시 연령, 백신의 용량, 백신접종후 시간경과 및 검사횟수 등에 영향을 받는 것으로 알려져 있다. 저자들은 BCG 접종후 시간경과, 재태기간, 성별 및 수유방법에 따라 Mantoux 검사의 반옹정도와 양전율에 차이가 있는지를 알아보기 위하여 이 연구를 시행하였다. 방 법 : 1994년 7월부터 1996년 6월까지 강남성모병원에서 출생한 신생아 85명(만삭분만아 71명, 미숙분만아 14명)을 대상으로 하였다. BCG백신은 국립보건원에서 제조한 것으로 생후 4주 이내에 0.05ml를 피내주사하였고, 접종후 3개월, 9개월, 15개월에 Mantoux 검사를 시행하였다. 검사시약은 PPD(5TU)를 사용하였다. BCG백신 접종과 Mantoux 검사의 결과 판독은 모두 동일인에 의하여 시행되었다. 결 과: 1) BCG 접종후 3개월, 9개월 및 IS개월에 Mantoux 검사에서 양전을은 만삭분만군 26.8%, 53.5%, 및 60.6%이었으며, 미숙분만군 14.3%, 28.6%, 50%로 만삭분만군에 비하여 미숙분만군에서 전반적으로 낮은 경향을 보였고 생후 3개월에 유의한 차를 나타내었다. 2) BCG 접종후 3개월, 9개월 및 15개월에 시행한 Mantoux 검사에서 경결의 평균 크기는 만삭분만군 7.2㎜, 8.5㎜, 9.4㎜를, 미숙분만군 5.0㎜ 6.6㎜, 8.2㎜를 보였으며 3개월에 검사한 경결의 크기는 두 군간에 통계학적으로 유의한 차가 있었다(P<0.05). 3) 만삭분만군 및 미숙분만군 모두에서 성별 및 수유방법에 따른 Mantoux 검사의 양전율은 유의 한 차가 없었다. 4) BCG백신의 합병증은 만삭분만군에서 9.9%로 림프절염, 지축적 궤양 및 열성 농양의 순이었으며 미숙분만군에서는 합병증이 한 예도 없었다. 결 론 : 이상의 결과에서 Mantoux 검사의 반응은 시간이 경과함에 따라 경결의 크기가 증가하고 양전율이 증가함을 알 수 있었으며 3개월 Mantoux 검사에서 미숙분만군에서 경결의 크기가 만삭분만군보다 작은 것은 미숙분만아의 면역학적 미성숙과 관련이 있을 것으로 생각되었다. Purpose : During the childhood, Mantoux test is very popular for detecting tuberculous infection and also useful for evaluating the immunity against tuberculosis after BCG vaccination. But the response and conversion rate is affected by the age of vaccination, the quality of vaccine, the duration after vaccination and the frequency of the test. This study was performed to investigate the difference of the response and conversion rate, according to duration after vaccination, gestational age, sex and feeding type. Methods : The subject was included 85 of infants (71 of full term infants, 14 of preterm infants) and was followed up 3 months, 9 months, 15 months after vaccination, from July, 1992 to June, 1994 in Kangnam St. Mary`s Hospital. We injected intradermally the BCG vaccine (0.05m1) which was produced by National Institute of Health and PPD (5TU) was used as a testing material of Mantoux test. Injecting BCG and interpreting the result of the Mantoux test was done by the same person. Results : 1) The positive conversion rates of Mantoux test which was done 3 month, 9 month and 15 month after vaccination were 26.8%, 53.5% and 60.6% in the full-term infant group, and 14.3%, 28.6%, and 50% in the preterm infant group. The conversion rate was significantly lower in preterm infant group as compared with that of the full-term infant group 3 months after vaccination. 2) The sizes of induration in Mantoux test, which was done 3 month, 9 month and 15 month after vaccination, are 7.2㎜, 8.5㎜, 9.4㎜ in the full-term infant group and 5.0mm, 6.6mm, 8.2mm in the preterm infant group. The size of induration was significantly lower in the preterm infant group as compared with that of the full-term infant group. 3) There is no difference in conversion rate of Mantoux test according to the sex, feeding type in both groups. 4) Complication rate for BCG vaccine is 9.9%, which is leaded by lymphadenitis, persistent ulcer, and febrile abscess as the order, in the full-term infant group and there are no complications in the preterm infant group. Conclusions : In those results, the size of induration and conversion rate is increased as the duration after vaccination, then lower conversion rate in the preterm infant group 3 months after vaccination may be due to immunological immaturity of preterrn infant.

Inhibitory Effect of BCG Cell-Wall Skeletons (BCG-CWS) Emulsified in Squalane on Tumor Growth and Metastasis in Mice

Yoo, Yung-Choon,Hata, Katsusuke,Lee, Kyung-Bok,Azuma, Ichiro The Pharmaceutical Society of Korea 2002 Archives of Pharmacal Research Vol.25 No.4

The antimetastatic effect of BCG-CWS, which was emulsified in an oil-in-water form with either Drakeol 6VR mineral oil (BCG-CWS/DK) or squalane (BCG-CWS/SQA), on lung metastasis produced by highly metastatic murine tumor cells, Colon26-M3.1 carcinoma cells and B16-BL6 melanoma cells, was investigated in syngeneic mice. An intravenous (i.v.) administration of BCG-CWS (100 mg/mouse) 1 day after tumor inoculation significantly inhibited tumor metastasis of both Colon26-M3.1 carcinoma and B16-BL6 melanoma cells in experimental lung metastasis models. No differences in the antitumor activity of the two oil-based formulations (BCG-CWS/DK and BCG-CWS/SQA) were obverved. However, BCG-CWS/SQA administered through subcutaneous (s.c.) route was shown to be effective only when it was consecutively injected (3 times) after tumor inoculation. An in vivo analysis for tumor-induced angiogenesis shwed that a single i.v. administration of BCG-CWS/SQA inhibited the number of tumor-induced blood vessels and suppressed tumor growth. Furthermore, the multiple administration of BCG-CWS/SQA given at on week intervals led to a significant reduction in spontaneous lung metastasis of B16-BL6 melanoma cells in a spontaneous metastasis model. These results suggest that BCG-CWS emulsified with squalane is a potent inhibitory agent of lung metastasis, and that the anti metastatic effect of BCG-CWS is related to the suppression of tumor growth and the inhibition of tumor-induced angiogenesis.

Effect of Maternal Immune Status on Responsiveness of Bacillus Calmette-Gurin Vaccination in Mouse Neonates

Choi, Jong Su,Kim, Ryang Yeo,Rho, Semi,Ewann, Fanny,Mielcarek, Nathalie,Song, Man Ki,Czerkinsky, Cecil,Kim, Jae-Ouk Korea Centers for Disease Control and Prevention 2012 Osong Public Health and Research Persptectives Vol.3 No.2

<P><B>Objectives</B></P><P>Bacillus Calmette-Guérin (BCG) vaccination has proven to be efficient in immunologically naïve infants; however, it has not been investigated that maternal natural exposure to <I>Mycobacterium</I> and/or BCG vaccine could influence the characteristics of immune responses to BCG in newborns. In this study, we analyzed whether the maternal immune status to <I>M tuberculosis </I>(<I>M tb</I>) can affect neonatal immunity to BCG using a mouse model.</P><P><B>Methods</B></P><P>Neonates were obtained from mice that were previously exposed to live BCG, to live <I>M avium</I>, or to heat-killed <I>M tb</I> H37Rv, and from naïve control mothers. One week after birth, the neonates were divided into two subgroups: one group immunized with live BCG via the subcutaneous route and the other group of neonates sham-treated. Interferon-gamma (IFNγ) secretion in response to <I>in vitro </I>stimulation with heat-killed BCG or purified protein derivative (PPD) was examined. Protection against <I>M tb</I> infection was evaluated by challenging mice nasally with live <I>M tb</I> H37Rv followed by counting colonies from spleen and lung homogenates.</P><P><B>Results</B></P><P>BCG-immunized neonates showed increased IFNγ secretion in response to heat-killed BCG or PPD. All mice in BCG-immunized neonates subgroups showed reduced bacterial burden (colony forming unit) in the lungs when compared with control naive neonate mice. However, no statistically significant difference was observed when comparing BCG-immunized mice born from mothers previously exposed to <I>M avium</I> or immunized with either heat-killed H37Rv or live BCG and mice born from naïve mothers.</P><P><B>Conclusion</B></P><P>The maternal immune status to <I>M tb</I> does not appear to impact on the immunogenicity of BCG vaccine in their progeny in our experimental conditions</P>

개원가의 신생아 BCG 접종 후 투베르쿨린 반응 평가 -접종 방법과 PPD 종류에 의한 차이 비교-

김완주,이선호,안상윤,양승재,오성희,Kim, Wan Ju,Lee, Sun Ho,Ahn, Sang Yoon,Yang, Seung Jae,Oh, Sung Hee 대한소아감염학회 2002 Pediatric Infection and Vaccine Vol.9 No.2

목 적 : BCG 접종 후 투베르쿨린 반응은 사용한 백신의 종류, 백신의 상태, 투여방법, 접종시 연령, 접종후 검사하기까지의 시간 등의 다양한 요인에 따라 다르게 나타난다. 최근 흉터가 작고 이상반응이 적은 경피용 BCG의 접종이 증가하고 있으나 투베르쿨린 반응 검사 및 예방 효과 등에 대한 자료가 거의 없는 실정이다. 본 연구는 개원가에서 실시한 BCG 및 투베르쿨린 반응을 평가하고 피내 BCG 백신과 경피용 BCG 백신과의 비교, 5TU PPD와 2TU PPD의 비교를 실시하였다. 방 법 : 1997년 2월부터 2001년 6월까지 세군데의 개인 소아과 의원에서 생후 1개월경 BCG 접종을 받은 후 투베르쿨린 반응을 실시한 200명의 소아를 대상으로 의무기록지를 이용한 후향성 조사를 실시하였다. 이 중 151명은 Tokyo 172주로 경피 접종하였고 49명은 피내 접종(28명:copenhagen주, 1명:French 1173P2주, 20명:미상)하였다. 경피 접종한 151명 중 129명(64.5%)은 2TU, 22명(11%)은 5TU PPD를 사용하였고 피내 접종한 49명 중 35명(17.5%)은 2TU, 14명(7%)은 5TU PPD를 사용하였다. 결 과 : 경피용 BCG 접종군에서 2TU PPD와 5TU PPD에 대한 경결의 크기(양성률)는 $9.2{\pm}4.4mm$(86.8%), $12.4{\pm}3.5mm$(95.5%)로 5TU PPD로 검사한 경우가 2TU PPD로 검사한 경우보다 통계적으로 유의하게 더 크게 나타났다(P=0.02). 피내용 BCG 접종군에서 2TU PPD와 5TU PPD에 대한 경결의 크기(양성률)는 각각 $5.7{\pm}5.1mm$(60%), $6.6{\pm}4.8mm$(71.4%)로 두 값 사이에 통계적으로 유의한 차이가 없었다(P=0.569). 2TU PPD에 대한 반응은 경피용 BCG 접종군에서의 경결 크기(양성률)가 $9.2{\pm}4.4mm$(86.8%), 피내용 BCG 접종군에서는 $5.7{\pm}5.1mm$(60%)로 경피용 BCG 접종군에서의 경결크기가 피내용 BCG 접종군에 비해 통계적으로 유의하게 크게 나타났다(P<0.001). 5TU PPD에 대한 반응은 경피용 BCG 접종군에서의 경결크기(양성률)는 $12.4{\pm}3.5mm$(95.5%), 피내용 BCG 접종군에서는 $6.6{\pm}4.8mm$(71.4%)로 역시 경피용 BCG 접종군에서 경결크기가 피내용 BCG 접종군에 비해 통계적으로 유의하게 크게 나타났다(P<0.001). 결 론 : 경피용 BCG(Tokyo 172균주) 접종군의 2TU PPD와 5TU PPD에 대한 반응은 피내용 BCG 접종군에 비해 경결의 크기가 유의하게 컸고, 경피용 BCG 접종군에서 5TU PPD의 경결 크기가 2TU PPD에 비해 유의하게 크게 나타났으므로 경피용 BCG 환아들에게서의 결핵 반응 검사 결과 판독의 새로운 기준이 수립되어야 할 것으로 사료된다. Purpose : The number of newborns vaccinated with BCG of Tokyo 172 strain, which has been claimed to cause lesser degree of local adverse events including scar, has recently been increasing. However, tuberculin response to this vaccine has inadequately been studied, especially with newborns cared at primary care clinics. We, therefore, performed a study in newborns vaccinated with BCG at private pediatric offices and evaluated the response to PPD 2TU or PPD 5TU following vaccination with percutaneous or intradermal BCG. Methods : Two hundred infants who had been cared at three private pediatric offices were retrospectively enrolled in the study. One hundred fifty one infants had received percutaneous BCG(Tokyo strain); 129 infants had had tuberuclin test with PPD 2TU and the rest of 22 infants with PPD 5TU. Forty nine infants had received intradermal BCG(28 infants Copenhagen strain, I infant French strain, 20 infants unknown); 35 infants had had tuberculin test with PPD 2TU, 14 infants(11%) with PPD 5TU. Results : In infants vaccinated with percutaneous BCG, the mean induration diameter in tuberculin test was significantly greater with PPD 5TU($12.4{\pm}3.5mm$) compared to PPD 2TU ($9.2{\pm}4.4mm$). In infants vaccinated with intradermal BCG, the mean induration diameters in tuberculin test were $5.7{\pm}5.1mm$ to PPD 2TU and $6.6{\pm}4.8mm$ to PPD 5TU, which were not significantly different. The tuberculin response to PPD 2TU was significantly greater in infants vaccinated with percutaneous BCG compared to those with intradermal BCG. The tuberculin response to PPD 5TU was also significantly greater in infants vaccinated with percutaneous BCG compared to those with intradermal BCG. Conclusion : Percutaneous BCG(Tokyo strain) seems to cause greater response to tuberculin compared to intradermal BCG and PPD 2TU induces weaker response compared to PPD 5TU. Acknowledging some discrepancies from the previously reported data, which might have been due to the different source of the study subjects, more studies are needed to establish the range of tuberculin response following BCG vaccination in order to differentiate from tuberculosis.

Studies on the prevention of tuberculosis in pet dogs 1. The effects of BCG pretreatment in pet dogs inoculated experimentally with Mycobacterium bovis

강종구,김창기,Kang, Jong-koo,Kim, Chang-ki The Korean Society of Veterinary Science 1992 大韓獸醫學會誌 Vol.32 No.1

Mycobacterium (M) bovis 를 인공감염시킨 개에 있어서 BCG의 전처치효과를 in vivo 및 in vitro에서 검토하였다. 개들은 BCG 전처치군, M bovis 단독처치군, 비감염대조군의 세군으로 나누었다. BCG는 M bovis 복강접종 3주일전에 0.2ml를 피내접종하였다. 결핵균 투여 4개월후에 전군을 도살하여 실험에 사용하였다. 도살시 모든 처치군에서 감염이 확인되었다. 병리조직학적으로 BCG전처치군의 폐장내에서는 경도의 macrophage의 침윤과 소상의 육아종 형성이 관찰되였으나 M bovis 단독처치군에 있어서는 보다 고도의 macrophage의 침윤, 중등도의 호중구의 침윤 및 중등도의 육아종의 형성이 확인되었다. 각 동물의 기관지폐포세정액을 분리하여 그 속의 총세포수와 각 세포의 분획을 검토하였다. 비감염 대조군의 기관지폐포세정액내의 총세포수는 두 처치군보다 훨씬 낮았으며 M bovis 단독처치군의 총세포수는 BCG 전처치군보다 1.8배 높았다. 이 세정액으로부터 폐포 macrophage를 분리배양하여 macrophage의 활성능과 결핵균의 증식능을 관찰하였다. BCG처치군은 M bovis 단독처치군에 비하여 높은 Fc receptor 활성(rosette 형성능, 탐식능)과 낮은 결핵균의 증식이 관찰되었다. 그러나 BCG의 전처치는 결핵균을 killing하지는 못하였다. 개에게 BCG를 전처치하면 폐내에 극소수의 결핵균이 지속적으로 잔존하지만 폐포 macrophage는 이미 항결핵성면역능을 지닌채로 계속 활성화된 상태로 존재하기 때문에 결핵에 대하여 예방효과를 갖는다고 사료된다. Dogs were divided into 3 groups of two each; Bacillie Calmette-Guerin(BCG) pretreatment, M bovis only treatment and uninfected control group. BCG were vaccinated intradermally with 0.2ml before 3weeks of M bovis intraperitoneal infection. Infection at necropsy 4months later was readily in the both treated dogs. Histopathologically, the BCG pretreated dogs produce the moderate accumulation of macrophages and focal granuloma formation in the lung, whereas the M bovis only treared dogs produce the accumulation of predominantly macrophages, occasionaly polymorphonuclear cells and the more larger granuloma Bronchoalveolar lavage(BAL) was obtained and total and differential cell counts were examined. Total number of BAL cells harvested from uninfected dogs is lower compared with those of the both treated groups. The total cell number of M bovis only treated dogs were singificantly higher 1.8 times than that of the BCG pretreated dogs. The Fe receptor activity and the growth of organism in alveolar macrophages obtained from BCG pretreated dogs were compared with that in macrophages from M bovis only treated dogs. BCG vaccination resulted in substantial macrophage activation, measured as increased Fc receptor mediated phagocytosis and rosette formation, as wells as the inhibition of intracellular mycobacteria multiplication. However, actibated macrophages taken from BCG pretreated dogs are incapable of killing the M bovis. Thus, these results suggest that BCG pretrearment in the dog may produce a protective effect against tuberculosis because active alveolar macrophages have acquired antituberculous immunity, although few mycobacteria within the lung remain in a metabolically active state.