OBJECTIVES: To circumvent the limitations of the current golden standard method, colony-forming unit (CFU) assay, for viability of Bacille Calmette–Guérin (BCG) vaccines, we developed a new method to rapidly and accurately determine the potency of BCG vaccines. METHODS: Based on flow cytometry (FACS) and fluorescein diacetate (FDA) as the most appropriate fluorescent staining reagent, 17 lots of BCG vaccines for percutaneous administration and 5 lots of BCG vaccines for intradermal administration were analyzed in this study. The percentage of viable cells measured by flow cytometry along with the total number of organisms in BCG vaccines, as determined on a cell counter, was used to quantify the number of viable cells. RESULTS: Pearson correlation coefficients of FACS and CFU assays for percutaneous and intradermal BCG vaccines were 0.6962 and 0.7428, respectively, indicating a high correlation. The coefficient of variation value of the FACS assay was less than 7%, which was 11 times lower than that of the CFU assay. CONCLUSION: This study contributes to the evaluation of new potency test method for FACS-based determination of viable cells in BCG vaccines. Accordingly, quality control of BCG vaccines can be significantly improved.
Administration, Cutaneous ; BCG Vaccine ; Cell Count ; Flow Cytometry* ; Fluorescein ; Methods ; Mycobacterium bovis ; Quality Control ; Stem Cells ; Vaccine Potency ; Vaccines*

Intravesical instillation of Mycobacterium bovis bacille Calmette–Guérin (BCG) has been used for treating nonmuscle invasive bladder cancer as the forefront of immunotherapy, but BCG is ineffective in approximately 30–40% of cases and disease recurs in up to 50% of patients. Recently BCG is considered an effective vehicle for delivery of antigens due to its unique characteristics, and the genetic control of these mycobacteria is advanced in the search for less toxic and more potent therapeutic agents for bladder cancer immunotherapy. We will discuss current advances in recombinant BCG construction, research, and future directions.
Administration, Intravesical ; BCG Vaccine ; Humans ; Immunotherapy* ; Mycobacterium bovis ; Urinary Bladder Neoplasms* ; Urinary Bladder*

PURPOSE: We compared prophylactic effects and complications of intravesical instillation of the Connaught and Tice strains bacillus Calmette-Guerin (BCG) in patients with stage pT1 bladder cancer. MATERIALS AND METHODS: A total of 98 patients with stage pT1 bladder cancer were treated with transurethral resection (TUR) between January 1992 and April 1998. Of the 98 patients, 51 received the Connaught strain BCG (81mg), 27 the Tice strain BCG (12.7mg) and 20 patients underwent TUR alone. The patients were followed-up for 18-78 months (mean 42.5months). The recurrence and progression rates, mean months to tumor recurrence, recurrence free survival rate, using Kaplan-Meier curve, and complications, were compared between the two BCG strain groups. RESULTS: The overall recurrence rate was 27.5% in the Connaught strain BCG group, 29.6% in Tice strain BCG group and 65% in TUR alone group. The mean months to tumor recurrence, and the recurrence free survival rate, showed that both BCG strain drugs were superior to TUR alone. Although the prophylactic efficacy of the Connaught strain BCG was a little higher than that of the Tice strain BCG, there were no significant differences in the recurrence rates and recurrence free survival rates between the two drugs. The incidences of complications were 94.1 and 85.2% in the Connaught strain BCG and Tice strain BCG groups, respectively. CONCLUSIONS: Both the Connaught and Tice BCG strains were superior to TUR alone in the prophylaxis of the recurrence in stage pT1 bladder cancer. There was no significant difference in the complication rates between the two groups. Therefore, both the Connaught and Tice BCG strains may be regarded as alternative treatments.
Administration, Intravesical ; Bacillus* ; BCG Vaccine ; Humans ; Incidence ; Mycobacterium bovis ; Recurrence ; Survival Rate ; Urinary Bladder Neoplasms* ; Urinary Bladder*

OBJECTIVETo observe the efficacy of local injection of bacillus calmette-guerin polysaccharide-nuclear acid (BCG-PSN) following CO2 laser resection on condyloma acuminatum (CA).

METHODSOne hundred and six patients with CA were randomized into BCG-PSN group (60 cases) and interferon group (46 cases), treated respectively with BCG-PSN local injection and interferon subcutaneous injection following CO2 laser resection and followed up for 3 months.

RESULTSThe healing rate of the BCG-PSN group was 88.3% and that of the interferon group was 71.7%.

CONCLUSIONThe BCG-PSN local injection following CO2 laser resection is effective for CA patients.

Adjuvants, Immunologic ; administration & dosage ; Adult ; Aged ; BCG Vaccine ; administration & dosage ; Condylomata Acuminata ; therapy ; Female ; Humans ; Laser Therapy ; Male ; Middle Aged ; Nucleic Acids ; administration & dosage ; Polysaccharides ; administration & dosage

The anti-tuberculosis Bacille de Calmette et Guerin (BCG) vaccine was developed between 1905 and 1921 at Pasteur Institutes of Lille in France, and was adopted by many countries. BCG strains comprise natural mutants of major virulence factors of Mycobacterium tuberculosis and that BCG sub-strains differ markedly in virulence levels. The tuberculosis became endemic in Korea after the Korean War (1950s). The BCG strain, which was donated by Pasteur Institutes, was brought to Korea in 1955, and the first domestic BCG vaccine was produced by the National Defense Research Institute (NDRI), current Korea Centers for Disease Control and Prevention (KCDC), in 1960. Since 1987, BCG manufacture work was handed over to the Korean Institute of Tuberculosis (KIT), the freeze-dried BCG vaccine was manufactured at a scale required to meet the whole amount of domestic consumption. However, since 2006, the manufacture of BCG vaccine suspended and the whole amount of BCG was imported at this point of time. Now KIT is planning to re-produce the BCG vaccine in Korea under the supervision of KCDC, this will be render great role to National Tuberculosis Control Program (NTP) and provide initiating step for developing new tuberculosis vaccines in Korea.
Academies and Institutes ; BCG Vaccine ; Centers for Disease Control and Prevention (U.S.) ; France ; Hand ; Korea ; Korean War ; Mycobacterium bovis ; Mycobacterium tuberculosis ; Organization and Administration ; Sprains and Strains ; Tuberculosis ; Tuberculosis Vaccines ; Virulence Factors

OBJECTIVETo probe into immunological mechanisms and clinical therapeutic effect of acupoint-injection of BCG polysaccharide nuclear acid (BCG-PSN) for treatment of condyloma acuminatum (CA).

METHODSTwo hundred cases were randomly divided into 4 groups. After removed the CA by laser, the treatment group (group A) was treated with acupoint-injection of BCG-PSN, the control group I (group B) with intramuscular injection of BCG-PSN, the control group II (group C) with intramuscular injection of interferon, and the blank control group (group D) with no treatment. The levels of cellular immune function were detected before treatment and after treatment of 6 months, and the cases of relapse were recorded.

RESULTSThe cured rate of 94.3% in the group A was significantly higher than 78.0% in the group B, 80.4% in the group C and 78.2% in the group D, with significant differences (P < 0.05); in the group A, CD4+ percent increased, CD8+ percent decreased, CD4+ /CD+ ratio increased, and NK cell activity increased with a low relapse rate, and with significant differences as compared with the control groups (P < 0.05, P < 0.01).

CONCLUSIONAcupoint-injection of BCG-PSN has a better therapeutic effect and it can obviously reduce the recurrence rate of CA. The cellular immunoregulatory action is one of the mechanisms of this therapy in preventing relapse of CA.

Acupuncture Points ; Adolescent ; Adult ; BCG Vaccine ; administration & dosage ; CD4-CD8 Ratio ; Condylomata Acuminata ; immunology ; therapy ; Female ; Humans ; Injections ; Killer Cells, Natural ; immunology ; Male ; Middle Aged ; Nucleic Acids ; administration & dosage ; Polysaccharides, Bacterial ; administration & dosage

Bacillus Calmette-Guerin (BCG) has been traditionally used as a vaccine against tuberculosis. Further, intravesical administration of BCG has been shown to be effective in treating bladder cancer. Although BCG contains a live attenuated strain of Mycobacterium bovis, complications such as M. bovis BCG infection caused by BCG administration are extremely rare. Here, we report a case of BCG infection occurring after intravesical BCG therapy. A 67-yr-old man presented with azotemia and weight loss. He had been diagnosed with bladder cancer 4 yr back, and had undergone transurethral resection of the bladder tumor and intravesical BCG (Tice strain) therapy at that time. An acid-fast bacterial strain was isolated from his urine sample. We did not detect Mycobacterium tuberculosis protein 64 (MPT-64) antigen in the isolates obtained from his sample, and multiplex PCR and PCR-reverse blot hybridization assay indicated that the isolate was a member of the M. tuberculosis complex, but was not M. tuberculosis. Finally, sequence analysis of 16S ribosomal RNA and DNA gyrase, subunit B (gyrB) suggested that the organism was M. bovis or M. bovis BCG. Although we could not confirm that M. bovis BCG was the causative agent, the results of the 3 molecular methods and the MPT-64 antigen assay suggest this finding. This is an important finding, especially because M. bovis BCG cannot be identified using common commercial molecular genetics tools.
Administration, Intravesical ; Aged ; BCG Vaccine/administration & dosage/*adverse effects ; DNA Gyrase/genetics ; Humans ; Male ; Mycobacterium Infections/*diagnosis/etiology ; Mycobacterium bovis/genetics/*isolation & purification ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Urinary Bladder Neoplasms/*therapy

Intravesical bacillus Calmette-Guerin (BCG) administration has been used as an adjuvant therapy after transurethral resection for superficial bladder cancer, but the exact mechanisms of its antitumor activity are not yet known. The aim of this study was to characterize the immunologic aspects of antitumor activity of BCG using an animal model. C3H/He inbred mice and murine bladder tumor cell line, MBT-2 were used. The changes in immune cells such as helper T cells, suppressor T cells, macrophages and natural killer cells in the bladder and spleen were analysed by immunohistochemical method in intravesical BCG instilled in normal bladder, MBT-2 implanted after electrocauterization of the bladder mucosa and MBT-2 implanted and intravesical BCG treated group. The changes in natural killer cell activity of the splenocytes and peritoneal lymphocytes were evaluated using 51chromium release assay at regular time intervals following intraperitoneal BCG instillation. The prophylactic anticancer effect was evaluated by observing the tumor growth in the intravesically BCG treated group after intravesical MBT-2 implantation. In immunohistochemical examination, a remarkable infiltration of macrophage and helper T cell was observed in the lamina propria of the bladder, and the helper and suppressor T cells ratio (Th/Ts ratio) was increased after intravesical BCG therapy. In 51chromium release assay, enhanced natural killer cell activity of the splenocytes and peritoneal lymphocytes was observed after intraperitoneal BCG inoculation. The growth of implanted tumor was suppressed following intravesical instillation of BCG. These results suggest that the antitumor activity of BCG is not related to the simple inflammatory reaction but to the local and systemic immune response in which helper T lymphocytes and mononuclear cells play an important role.
Animals ; BCG Vaccine/administration & dosage/*therapeutic use ; Cells, Cultured ; Female ; Killer Cells, Natural/immunology ; Mice ; Mice, Inbred C3H ; Spleen/pathology ; T-Lymphocytes/*immunology ; Urinary Bladder/pathology ; Urinary Bladder Neoplasms/immunology/pathology/*therapy

This study was performed to estimate the rate of boosted reaction in the two-step tuberculin skin test (TST) and to evaluate the associated factors among military personnel of South Korea, which has an intermediate burden of tuberculosis (TB) and a routine bacille Calmette-Guerin (BCG) vaccination policy. Two-step TST was performed on 264 military personnel who did not have a history of close contact to TB. Subjects with a negative reaction to the first test of <10 mm had a second TST applied 1 week later on the other forearm. A positive result (> or =10 mm) on the initial TST was observed in 126 (48%) of the subjects. A boosted reaction on the second TST developed in 32 (23%) of the 124 subjects with a negative initial TST. In multiple logistic regression analysis, the size of the initial TST reaction was the only factor associated with a boosted reaction on the second TST. The high rate of boosted reaction among healthy adults in South Korea suggests that two-step TST should be performed to assess the baseline TST reactivity in settings with an intermediate burden of TB and routine BCG vaccination policy, especially among subjects with an initial TST reaction that is > or =5 mm.
Adult ; BCG Vaccine/*administration & dosage ; Humans ; Hypersensitivity/*diagnosis ; Incidence ; Korea/epidemiology ; Logistic Models ; Male ; Middle Aged ; *Military Personnel/statistics & numerical data ; Prevalence ; *Tuberculin Test ; Tuberculosis, Pulmonary/*diagnosis/epidemiology/prevention & control

INTRODUCTION: Kawasaki disease (KD) is a challenging diagnosis. Erythema and induration of the Bacillus Calmette-Guérin (BCG) site is increasingly recognised as a significant clinical clue. However, there is little data to support its specificity for KD as compared to other febrile illnesses. We aimed to evaluate BCG reaction or induration as a diagnostic tool for KD. METHODS: A retrospective case-controlled study of patients discharged with a diagnosis of KD from 2007 to 2010 was conducted. Another group of patients admitted over the same period for possible KD, but later found not to have KD, served as control. RESULTS: Significantly more infants with KD (69.7%) had BCG site changes than older children (27.8%; p < 0.001). It also presented earlier in the course of KD; < 5 days (53.3%) compared to ≥ 5 days of fever (30.0%; p < 0.001). Positive predictive value of BCG site reaction or induration for KD was 90.8% (95% confidence interval [CI] 0.819-0.962) for infants and 96.2% (95% CI 0.868-0.995) for older children. The prevalence rate of changes at the BCG site was 9.9% among patients with non-KD febrile illnesses and 42.6% among patients with KD. CONCLUSION BCG site reaction or induration is a useful clinical clue for the diagnosis of KD in both infants and older children, with a higher prevalence in infants. Physicians should consider KD in children with febrile illness and redness or crust formation at the BCG site, especially in view of low rates of BCG reaction or induration in non-KD febrile illnesses.
BCG Vaccine ; administration & dosage ; adverse effects ; Case-Control Studies ; Child, Preschool ; Erythema ; complications ; epidemiology ; Female ; Fever ; complications ; Humans ; Infant ; Male ; Mucocutaneous Lymph Node Syndrome ; complications ; diagnosis ; epidemiology ; Risk Factors ; Singapore ; epidemiology