Asian Pacific Journal of Allergy and Immunology

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Development of flow cytometry for detection and quantitation of red cell bound immunoglobulin G in autoimmune hemolytic anemia with negative direct Coombs test.
(2011-12) Thedsawad, Anchalee; Taka, Orathai; Wanachiwanawin, Wanchai
About 2-10% of patients with warm-antibody autoimmune hemolytic anemia (WAIHA) exhibit a negative direct Coombs test (DAT), requiring more sensitive tests, including detection of RBC-bound immunoglobulins by flow cytometry, for diagnosis. In this study, the optimal conditions for detection and quantitation of RBC-bound IgG by flow cytometry were studied using blood samples from six patients with AIHA and two healthy individuals. Quantitation of RBC-bound IgG was performed using quantum simply cellular (QSC) beads coated with goat anti-mouse IgG antibodies. For detection of RBC bound IgG, a 60-minute incubation of all blood samples with 40 μl of 1:10 dilution of FITC-conjugated mouse anti-human IgG gave mean fluorescent intensity (MFI) values comparable to experiments using larger amounts or higher concentrations of the anti-human IgG. The acquired antibody binding capacity (ABC) values (or IgG molecules) for each QSC bead level, at 40 μl of 1:5 and 1:10 dilution of anti-human IgG for 60 minutes were close to the manufacturer-assigned ABC values. The IgG molecules per RBC in all six patients with positive DAT of 4+, 3+, 2+, 1+, trace and negative DAT were 31,725, 3,823, 1,753, 524, 260 and 88 respectively and in two healthy individuals with negative DAT they were 104 and 78.
A pediatric case of anaphylaxis due to octreotide.
(2011-12) Azkur, Dilek; Yoldas, Tamer; Toyran, Muge; Kocabas, Can Naci
Octreotide is an octapeptide that mimics natural somatostatin pharmacologically. It is a potent inhibitor of growth hormone, glucagon and insulin, which is used for treatment of acromegaly, symptomatic treatment of carsinoid tumours, and vasoactive intestinal peptide secreting tumors. It is also used for chylothorax, chemotherapy induced diarrhea and, as it inhibits the exocrine production of pancreatic enzymes, for acute and chronic pancreatitis. Gallbladder stones, diarrhea, nausea, vomiting, hypoglycemia/hyperglycemia, headache, and abdominal discomfort are some of the common adverse effects of octreotide and it may rarely cause anaphylaxis. We present here a child who had chronic pancreatitis and had an anaphylactic reaction to octreotide. To our knowledge this is the first pediatric case of anaphylaxis with octreotide who was successfully desensitized.
Omalizumab treatment in severe adult atopic dermatitis.
(2011-12) Thaiwat, Supitchaya; Sangasapaviliya, Atik
Atopic dermatitis (AD) is one of the most common chronic skin diseases. Treatment options include lubricants, antihistamines, and corticosteroids in either topical or oral forms. Severe AD is frequently recalcitrant to these medications. We reported three cases of severe AD patients who had elevated of IgE levels and failed to response to several prior medical treatment. After being treated with Omalizumab (humanized monoclonal anti-IgE antibody), the patients had marked alleviation of symptoms with improved Eczema Area and Severity Index (EASI) and pruritic scores. No patient experienced adverse effect.
The difference in IL-1β, MIP-1α, IL-8 and IL-18 production between the infection of PMA activated U937 cells with recombinant vaccinia viruses inserted 2004 H5N1 influenza HA genes and NS genes.
(2011-12) Vongsakul, Molvibha; Kasisith, Jitra; Noisumdaeng, Pirom; Puthavathana, Pilaipan
Background: The severity of avian influenza H5N1 disease is correlated with the ability of the virus to induce an over production of pro-inflammatory cytokines from innate immune cells. However, the role of each virus gene is unknown. To elaborate the function of each virus gene, the recombinant vaccinia virus inserted HA and NS gene from the 2004 H5N1 virus were used in the study. Methods: U937 cells and PMA activated U937 cells were infected with recombinant vaccinia virus inserted with HA or NS gene. The expressions of HA and NS proteins in cells were detected on immunofluorescence stained slides using a confocal microscope. The cytokine productions in the cell supernatant were quantitated by ELISA. Results: The recombinant vaccinia virus inserted with HA genes induces the production of IL-1β, MIP-1α, IL-8 and IL-18 cytokines from PMA activated U937 cells significantly more than cells infected with wild type vaccinia, whereas the recombinant vaccinia virus inserted with NS genes it was similar to that with the wild type vaccinia virus. However, there was no synergistic nor antagonistic effect of HA genes and NS genes in relation to cytokines production. Conclusion: Only the HA gene from the 2004 H5N1 virus induces IL-1β, MIP-1α, IL-8 and IL-18 cytokine productions from activated U937 cells. The same HA gene effect may or may not be the same in respiratory epithelial cells and this needs to be explored.
Antiallergic effect of Ostericum koreanum root extract on ovalbumin-induced allergic rhinitis mouse model and mast cells.
(2011-12) Jung, Hyo Won; Jung, Jin Ki; Park, Yong Ki
Objective: In the present study, the anti-allergic effect of OR extract was evaluated on an ovalbumin (OVA)-induced allergic rhinitis in mice and rat peritoneal mast cells (RPMC). Methods: Balb/c mice were systemically sensitized to OVA followed by intraperitoneal and nasal allergen challenges. We investigated the effect of OR extract on allergic symptoms, serological marker production and histological changes of the nasal mucosa in a mouse model of allergic rhinitis. We observed mast cell degranulation and detected the production of histamine and inflammatory cytokines by ELISA. Results: Compared to the OVA-control group, oral administration of OR extract at doses of 50 and 100 mg/kg significantly (p <0.001) decreased the serum levels of histamine, OVA-specific IgE and Th2 cytokine, IL-4 as well as increasing Th1 cytokine, IFN-γ. Oral administration of OR extract also attenuated disease progression as determined by nasal symptoms and histological changes of the nasal mucosa in OVA-sensitized mice. Furthermore, treatment with OR extract at doses of 0.2, 0.5 ad 1 mg/mL in RPMC significantly (p <0.01, p <0.001 and p <0.001, respectively) decreased compound 48/80-induced histamine release and suppressed mast cell degranulation. Treatment with OR extract in RPMC also inhibited PMA/A23187-induced production of The mechanism of action underlying OR extract in allergic inflammation appears to be inhibition of the phosphorylation of ERK1/2 and p38 MAPK, in addition to blocking of the NFκB pathway. Conclusions: These results indicate that OR extract has the potential to be a source of anti-allergic agents for use in allergen and/or mast cell-mediated diseases including allergic rhinitis.
Immunoglobulin G (IgG) subclass deficiency in Thai children.
(2011-12) Visitsunthorn, Nualanong; Hengcrawit, Wiriya; Jirapongsananuruk, Orathai; Luangwedchakarn, Voravich
Background: Patients with Immunoglobulin G (IgG) subclass deficiency may suffer from recurrent infections, mainly sino-pulmonary infection. Objective: To determine the epidemiology of IgG subclass deficiency in Thai children at a tertiary care hospital and to compare the differences between children who were diagnosed with IgG subclass deficiency by using low level criteria [less than 2 standard deviation (SD) of normal levels for age] and by using low percentage criteria (proportion of each IgG subclasses/total IgG). Methods: The study was a descriptive study of 55 children up to 15 years old with recurrent infections diagnosed as having IgG subclass deficiency but no acquired or other primary immune deficiencies except for IgA and/or IgM deficiency. Result: Isolated IgG3 subclass deficiency was the most common IgG subclass deficiency (56.4%). IgG3 subclass deficiency, either isolated or combined with other IgG subclass deficiency, was found in 85.5% of the cases. The common age of onset was between birth and five years of age. The most common presenting symptom was recurrent sinusitis (83.6%). Majority of the cases (89.3%) were diagnosed by low percentage criteria while 12.7% were diagnosed by low level criteria. All cases with low levels of IgG subclass antibodies also had low percentages. There were no statistically significant differences in the clinical manifestations and management methods between the children who were diagnosed by low level and low percentage. Conclusion: IgG3 subclass deficiency was the most common IgG subclass deficiency in Thai children. The most common presenting symptom was recurrent sinusitis. Although the diagnosis could be made in the patients with recurrent upper respiratory infection by using low level criteria, but the diagnosis should be considered when the low percentage criteria are met.
Type I hereditary angioedema in Taiwan – clinical, biological features and genetic study.
(2011-12) Lei, Wei Te; Shyur, Shyh-Dar; Huang, Li-Hsin; Kao, Yu-Hsuan; Lo, Chia-Yi
Background: Hereditary angioedema (HAE) is a rare, autosomal dominant inherited disease which is caused by a genetic deficiency of C1 esterase inhibitor (C1 INH). There have only been a few case reports in Taiwan to date. Objective: To describe the clinical features of type I HAE in Taiwanese patients. Methods: Three unrelated Taiwanese families with type I HAE are reported, and one case of a family from a review of PubMed was reviewed. Clinical manifestations, diagnostic examinations, management and genetic studies were analyzed. Results: Including this report, 19 patients had low C1 INH and low C4 levels and were diagnosed with type I HAE. Only 11 (57.9%) patients were symptomatic. Recurrent skin swelling and edema over the four extremities or trunk were reported in all symptomatic patients (100%). 45.5% of the patients recalled laryngeal attacks and one patient died from asphyxia. 18.2% of the patients experienced abdominal symptoms. The age at the beginning of clinical symptoms ranged from 5 to 30 years (mean ± SD: 20.82 ± 7.88 years). The diagnosis tended to be delayed (range from 1 to 39 years; mean ± SD: 8.45 ± 11.04 years). Nine patients had a mutant C1 INH gene, and two patients received long-term prophylaxis with danazol. Conclusion: The prevalence of hereditary angioedema in Taiwan is low. Persons with low levels of C1 INH who were clinically symptomatic accounted for only 57.9% of the cases in our study, which is far lower than previous reports from other countries. Ethnic differences may be the reason for this finding. Further genomic studies are needed to elucidate the genetic penetrance of C1 INH deficiency in Taiwan.
Prevalence and clinical characteristics of adult-onset atopic dermatitis with positive skin prick testing to mites.
(2011-12) Kulthanan, Kanokvalai; Chularojanamontri, Leena; Manapajon, Araya; Nuchkull, Piyavadee
Background The clinical role of house dust mite (HDM) in atopic dermatitis (AD) is still controversial. Objective The aim of the study is to assess the prevalence, clinical relevance and characteristics of adult-onset AD patients with positive skin prick tests (SPT) to mites. Methods The case record forms of adult-onset AD patients who underwent SPT at the Skin Allergy Clinic, Siriraj Hospital were reviewed. Results Forty-one of 62 patients (66.1%) had positive SPT to mites. The frequency of intrinsic AD among adult-onset AD was 4.8% (3/62). SPT to HDM tended to be positive in patients who had personal or family history of atopy, positive SPT to several specific antigens or who presented with elevated serum IgE, chelitis, recurrent conjunctivitis and perifollicular accentuation, respectively. Conclusion The prevalence of adult-onset AD patients with mite sensitivity was high. There were some notable features that tended to be present in mite sensitive adult-onset AD patients.
Predictors of childhood food allergy: significance and implications.
(2011-12) Al-Hammadi, Suleiman; Zoubeidi, Taoufik; Al-Maskari, Fatma
Background: Food allergy is common in children, and its occurrence is strongly associated with other allergies including anaphylaxis. Both genetic (e.g., CD14, STAT6, IL-10, SPINK5, and FOXP3 genes) and environmental (e.g., early exposure to highly allergic food) factors appear to contribute to food allergy. Method: Cross-sectional study involved children in public primary schools in Al-Ain city (United Arab Emirates). 660 students from the chosen classes were provided with 35 questions to be answered by their parents with a response rate 60.2%. The objective of the study was to determine predictors for food allergy in children. Result: Significant associations were found between childhood food allergy and a history of personal allergy (atopic dermatitis, asthma or allergic rhino-conjunctivitis) or immediate family members with food allergy or other allergic diseases. The best predictors for childhood food allergy were a personal history of asthma (p <0.001), a personal history of atopic dermatitis (p <0.001), a paternal history of atopic dermatitis (p =0.005) and a paternal history of allergic rhino-conjunctivitis (p =0.012). Discussion: These results are consistent with the notion that “various forms of allergy, including childhood food allergy are hereditarily coupled”. Thus, predicting childhood food allergy provides an opportunity to prevent or ameliorate the symptoms.
Clinical features of adult and pediatric anaphylaxis in Taiwan.
(2011-12) Hsin, Yi-Chen; Hsin, Yi-Chin; Huang, Jing-Long; Yeh, Kuo-Wei
Background: Most epidemiologic studies of anaphylaxis have been on Western populations, leaving the clinical and demographic pattern of this acute allergic condition in Asia unclear. Objective: To investigate the clinical characteristics of patients with anaphylaxis in Chang Gung Memorial Hospital, the largest medical center in Taiwan. Methods: We conducted a retrospective analysis of 201 patients who visited the emergency department or were admitted to the hospital for anaphylaxis from 2000 to 2010. We analyzed the causes, clinical presentation, and management, and also compared adult and pediatric cases. Results: The average patient age was 43.3 years. Mortality from anaphylaxis was 0.5% (1/201). The annual number of cases presenting with anaphylaxis increased throughout the decade we studied. Seven types of etiology were identified: medication (53%), contrast medium (24%), idiopathic condition (8%), food (5%), blood transfusion (4%), insect sting (3%), and others (3%). Skin and respiratory presentations are more common in children than in adults (skin presentation, 81% vs. 51%, p =0.002); respiratory presentation, 74% vs. 49%, p =0.011), and cardio-vascular presentation is more frequent in adults than in children (83% vs. 61%, p =0.006). Clinical presentations with angioedema, gastro-intestinal and neurological system involvement, and management were not significantly different between adults and children. Conclusions: We conclude that anaphylaxis in Taiwan appears to be increasing, just as in the West, but shows a different clinical picture; medication rather than food was the most common cause of anaphylaxis in our population. Moreover, food-induced anaphylaxis in children is not so prevalent in Taiwan.
Congestion and sleep impairment in allergic rhinitis.
(2011-12) Sardana, Niti; Craig, Timothy J
Allergic rhinitis is a disease with an increasing prevalence throughout the world that severely affects the quality of life of individuals suffering from it. Nasal congestion is the most common and bothersome symptom, and is often associated with sleep-disordered breathing, which is thought to be the reason for sleep impairment in individuals with rhinitis. The end result is a decrease in quality of life and productivity and an increase in daytime sleepiness. Treatment with intranasal corticosteroids has been shown to reduce nasal congestion. Data on sleep-related end points from clinical trials of intranasal corticosteroids indicate that this reduction is associated with improved sleep, reduced daytime fatigue, and improved quality of life. Other therapies, such as montelukast, also have a positive influence on congestion and sleep. This review examines nasal congestion and the associated sleep impairment of allergic rhinitis patients. It explores the adverse effects of disturbed sleep on quality of life and how these conditions can be reduced by therapies that decrease congestion.
Eulogy for Professor Chua Kaw Yan.
(2011-12) Bee, Wah Lee
Frequency of the HLA-B*1502 allele contributing to carbamazepine-induced hypersensitivity reactions in a cohort of Malaysian epilepsy patients.
(2011-09) Then, Sue-Mian; Rani, Zam Zureena Mohd; Raymond, Azman Ali; Ratnaningrum, Safrina; Jamal, Rahman
We describe the association of the HLA-B*1502 allele in 27 epilepsy patients (19 Malays, 8 Chinese) treated with carbamazepine (CBZ) at the UKM Medical Center (UKMMC), 6 with CBZ-Steven Johnson Syndrome (CBZ-SJS), 11 with CBZ-induced rash, 2 with suspected phenytoin-induced rash and 8 negative controls. Our study showed that 10 (6 Malay, 4 Chinese) patients were positive for HLA-B*1502. Out of the 10 patients, six were confirmed to have CBZ-SJS (p =0.0006), while four patients developed a skin rash. However there were 6 Malay patients and 1 Chinese patient that developed a skin rash after CBZ administration who were not positive for the allele, indicating that there might be more that one allele associated with CBZ-induced hypersensitivity. Another 2 patients were suspected of having phenytoin-induced rash, instead of CBZ, and these patients did not have HLA-B*1502. In conclusion, this study confirmed the association of HLA-B*1502 with CBZ-SJS among Malaysian epilepsy patients, however there might be other genes that could be responsible for the CBZ-induced rash.
Cutaneous adverse reactions to sulfonamide antibiotics.
(2011-09) Chantachaeng, Weeranut; Chularojanamontri, Leena; Kulthanan, Kanokvalai; Jongarearnprasert, Kowit; Dhana, Nareemon
Background: Sulfonamides are divided into two main groups which are sulfonamide antibiotics and sulfonamide non-antibiotics. The wide use of sulfonamide antibiotics leads to increasing incidence of sulfonamide cutaneous reactions. Objective: The purpose of this study is to explore the cutaneous manifestations induced by sulfonamide antibiotics in a large number of Thai patients, including human immunodeficiency virus (HIV) and non-HIV infected individuals. The second purpose is to determine the risk factors for development of sulfonamide cutaneous reactions. Methods: We retrospectively studied 191 patients with sulfonamide antibiotics cutaneous reactions attending the adverse drug reaction center, Siriraj Hospital, Bangkok between 2006 and 2010. Results: Majority of the patients was female (59.7%).Maculopapular rash was the most common cutaneous manifestation (37.7%), followed by fixed drug eruption (22%), angioedema with or without urticaria (12.6%) and urticaria alone (12%). Among those with known HIV serology, maculopapular eruption occurred more frequently in the HIV positive group while fixed drug eruption occurred more frequently in HIV-negative group. Conclusion: From our study, there were no significant determination factors to develop serious drug reactions. However, the HIV-positive status and lower level of CD4 count had a tendency to increase risk of developing serious cutaneous reactions.
Exercise-induced bronchoconstriction in rhinitis children without asthma.
(2011-09) Rakkhong, Kasalong; Kamchaisatian, Wasu; Vilaiyuk, Soamarat; Sasisakulporn, Cherapat; Teawsomboonkit, Wanlapa; Pornsuriyasak, Prapaporn; Benjaponpitak, Suwat
Background: Exercise-induced bronchoconstric-tion (EIB) is a condition that leads to limited participation in sports. Prevalence of EIB ranges from 5-20% in general population to 40 - 90% in asthmatic patients. But the prevalence among rhinitis children with EIB remains debatable. We aimed to determine the prevalence of EIB in non-asthmatic children with rhinitis. Methods: A cross-sectional study was performed on 53 rhinitis patients without known asthma who attended the Pediatric Allergy Clinic between March 2009 and February 2010.They all underwent physical examination, skin prick test, pulmonary function tests and an exercise challenge test (ECT) on a treadmill. A positive ECT was defined as a decrease in FEV1 > 10% after exercise. baseline pulmonary function and the decline in FEV1 after exercise. Results: The patients’ mean age was 12.3±2.6 years. Most of them (60.4%) had moderate to severe persistent rhinitis. Eleven out of 53 patients (20.7%) had EIB. The peak time for occurrence of EIB was 10 minutes after exercise. Most of the EIB episodes observed were of mild degree. Patients who had persistent symptoms and a short duration of rhinitis treatment prior to the ECT day had more positive ECT results (72.7% vs. 28.6%, p 0.013, 0.2 years vs.1.9 years, p 0.012, respectively). The history (Hx) was not a reliable means of identifying children who had EIB (ECT+/Hx+ 54.6%, ECT-/Hx+ 54.8%). There was no significant relationship between Conclusion: The prevalence of EIB in rhinitis children without asthma is 20.7%. History, physical examination and pulmonary function are insufficient to diagnose EIB.
Association of TGF-β1 , IL-4 and IL-13 gene polymerphisms with asthma in a Chinese population.
(2011-09) Yang, Xue-Xi; Li, Fen-Xia; Wu, Ying-Shong; Wu, Dong; Tan, Jia-Yu; Li, Ming
Background: Asthma is a common respiratory disease caused by genetic and environmental factors. It has been suggested that TGF-β1, IL-4 and IL-13 play important roles in asthma. Objectives: We attempted to confirm the roles of TGF-β1, IL-4 and IL-13 polymorphisms in asthma in a Chinese population. Methods: Five SNPs (rs1800469, rs2241712, rs2070874, rs20541 and rs1800925) in TGF-β1, IL-4 and IL-13 were genotyped using the MassArray SNP genotyping system. Allelic and genotypic associations between these SNPs and asthma were evaluated using logistic regression analysis. Results: The CT genotype of rs1800469 and T allele of rs20541 were significantly associated with asthma. Among atopic subjects, the CT genotype of rs1800469 and GA genotype of rs2241712 decreased the risk of asthma, while the CC genotype of rs2070874 showed a decreasing trend of asthma risk with a borderline significance. No significant association was found between rs1800925 and asthma. Conclusion: In the present study, we confirmed the association of rs1800469 in TGF-β1 and rs20541 in IL-13 with asthma and found a trend toward association between rs2241712 in TGF-β1 and rs2070874 in IL-4 with asthma among atopic subjects, suggesting TGF-β1, IL-4 and IL-13 may be associated with the susceptibility and development of asthma in this Chinese population.
Increased expression of aryl hydrocarbon receptor and interleukin 22 in patients with allergic asthma.
(2011-09) Zhu, Jing; Cao, Yong; Li, Kaiyan; Wang, Zhengyun; Zuo, Peng; Xiong, Weining; Xu, Yongjian; xiong, shengdao
Objectives: We sought to determine whether the aryl hydrocarbon receptor (AhR) and interleukin (IL)-22 may be involved in the pathogenesis of the peripheral blood mononuclear cells (PBMCs) in allergic asthmatic patients and whether their expression may be related to the severity of the disease. Methods: Blood samples were obtained from each subject with allergic asthma (n =18), controlled asthma (n =17) and healthy controls (n =12) respectively. The PBMCs were collected for AhR mRNA detection by real-time quantitative polymerase chain reaction (PCR). The plasma was collected for IL-22 protein detection by enzyme-linked immunosorbent assay (ELISA).Results: The expression of AhR mRNA in PBMCs and IL-22 protein in plasma of patients with allergic asthma were higher than those in controlled asthma cases and healthy controls. The plasma concentrations of IL-22 had negative correlation with the predicted percentage of forced expiratory volume in the first second (FEV1%) and the percentage of FEV1 and forced vital capacity (FEV1/FVC%) and it was positively correlated with the asthma severity score (ASS) of the asthmatics. Conclusion: Our results suggested that both AhR and IL-22 might be involved in the pathogenesis of allergic asthma in human and the level of IL-22 might have some relationship with the severity of the disease.
Clinical manifestations and BTK gene defect in 4 unrelated Taiwanese families with Bruton’s disease.
(2011-09) Lee, Kuo-Hsi; Shyur, Shyh-Dar; Chu, Szu-Hung; Huang, Li-Hsin; Kao, Yu-Hsuan; Lei, Wei-Te; Cheng, Chieh-Han; Lo, Chia-Yi; Chen, Chen-Kuan; Liu, Ling-Chun
Background and objective: X-linked agammaglo-bulinemia (XLA, also called Bruton’s disease) is is an X-linked recessive disorder characterized by recurrent bacterial infections, usually occurring in the first few years of life. Here, we report the results of a BTK gene mutation screening study that was performed in Taiwanese families with the BTK gene defect to further understand the inheritance patterns of XLA patients in Taiwan and to avoid new cases of XLA within families. Materials and methods: In this study, 52 members of 4 unrelated Taiwanese families with the BTK gene defect were enrolled. We studied the immunologic reports of 6 symptomatic living male patients with confirmed BTK gene defects and correlated the findings with their clinical symptoms. The genomic DNA of the subjects was subjected to direct sequencing mutation analysis. Results: We screened 52 members of 4 unrelated Taiwanese families with the BTK gene defect for BTK gene mutation and found that there were 6 symptomatic living patients with a confirmed defect, 7 symptomatic deceased patients highly suspected to have had the defect and 11 asymptomatic female carriers. Conclusions: This is the first report in a series of the thorough screening for the BTK mutation and its carrier status in 4 unrelated Taiwanese families. One pedigree of our study comprises 4 generations. A complete BTK gene mutation study for the patient’s family members is strongly suggested.
A single injection of 19 kDa carboxy-terminal fragment of Plasmodium yoelii merozoite surface protein 1 (PyMSP119) formulated with Montanide ISA and CpG ODN induces protective immune response in mice.
(2011-09) Hirunpetcharat, Chakrit; Mahakunkijcharoen, Yuvadee; Jeamwattanalert, Pimmada; Kittigul, Leera; Mahannop, Pakpimol; Pichyangkul, Sathit
Objective: To investigate the efficacy of a vaccine formulation of the 19 kDa conserved carboxyl-terminal fragment of Plasmodium yoelii merozoite surface protein-1 (PyMSP119) formulated with CpG ODN 1826 and Montanide ISA51 or ISA720 when used to immunize mice by a single injection. Methods: Groups of BALB/c mice were immunized parenterally with one, two or four injections with PBS or PyMSP119 formulated with CpG ODN in ISA51 or ISA720. Sera were collected weekly and assessed for total IgG and IgG subclass titers. Protection was tested by challenge infection with P. yoelii YM. Results: Interestingly, single injection immunization showed the same kinetics of antibody responses as two- or four-injection immunization. However, the peak antibody response induced by PyMSP119 in CpG ODN and ISA51 appeared earlier than that induced by PyMSP119 in CpG ODN and ISA720 (28 days vs 41 days). At day 63 after the first injection, the PyMSP119-specific IgG antibody levels by single injection and four- injection immunizations were not different. However, the levels of the IgG2a antibody subclass were significantly lower by single injection immunization with PyMSP119 in CpG ODN and ISA720. The antibodies were sustained at high levels for at least 20 weeks. After challenge infection, all mice immunized by a single injection of PyMSP119 in CpG ODN and ISA51 survived with low-grade parasitemia, while 50% of mice immunized with PyMSP119 in CpG ODN and ISA720 died with high levels of parasitemia. Conclusion: These findings suggest that MSP119 immunization by a single injection can induce protective immunity, particularly when formulated with an appropriate strong adjuvant.
Apoptosis, production of MMP9, VEGF, TNF- and intracellular growth of M. tuberculosis for different genotypes and different pks15/1 genes.
(2011-09) Yorsangsukkamol, Juthaporn; Chaiprasert, Angkana; Palaga, Tanapat; Prammananan, Therdsak; Faksri, Kiatichai; Palittapongarnpim, Prasit; Prayoonwiwat, Narapon
Background: A previous study of IS6110 RFLP and spoligotyping of M. tuberculosis isolates from 152 Thai patients with tuberculous meningitis revealed a significantly higher percentage (57%) of the Beijing genotype as compared to isolates obtained from pulmonary tuberculosis. We postulated that the M. tuberculosis Beijing genotype is likely to be more virulent than others. Objectives: Ten M. tuberculosis cerebrospinal fluid (CSF) isolates from five RFLP groups, together with different characteristics of pks15/1, M. tuberculosis H37Rv and M. bovis BCG, were investigated for their virulence in vitro. Methods: In this study, THP-1 cells were used as host cells to determine the intracellular growth and the induction of MMP9, VEGF, TNF-α and apoptosis. Determinations of the cytokine production and apoptosis were based on available commercial kits using ELISA techniques. Results: No significant difference in intracellular multiplication was found between the M. tuberculosis CSF isolates. Three isolates, consisting of 2 Nonthaburi and 1 heterogeneous isolate, were found to stimulate high TNF-α and MMP-9 production during the early infection period.They were isolated from 3 different patients, 2 of whom died with initial stages II and III. This result suggested that there might be an association between TNF-α and MMP-9 production that could account for the specific virulent nature of Nonthaburi strains. VEGF production was determined and comparable levels were found in all isolates. No significant apoptosis was detected in M. tuberculosis CSF isolates. No significant differences suggesting that the 2 Beijing strains are more virulent than the others were observed. Conclusion: The predominance of the Beijing strains in cases of tuberculous meningitis (TBM) in Thai patients is not a result of their hypervirulence.

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