scholarly journals Gene expression tryptophan aspartate coat protein in determining latent tuberculosis infection using immunocytochemistry and real time polimerase chain reaction

Author(s):  
Rebekah J. Setiabudi ◽  
Ni Made Mertaniasih ◽  
Muhammad Amin ◽  
Wayan Tunas Artama

Background: Tuberculosis (TB) remains a major cause of morbidity and mortality worldwide. Problem of Latent Tuberculosis Infection (LTBI) is increasing in number especially in countries with high TB incidence rate, such as Indonesia. Although not every LTBI will become active TB, if untreated and not handled appropriately it can still be a source of transmission and may increase the rate of resistance to the first-line TB drugs. Mycobacterium tuberculosis as a cause of tuberculosis disease is an intracellular pathogens that survives within the phagosome of host macrophages. Several host factors are involved in this process, including the Tryptophan Aspartate-containing Coat Protein (TACO). TACO is a protein recruited and retained by viable Mycobacterium tuberculosis on the surface of the phagosome membrane to maintain its survival in phagosome, because the presence of TACO plays an important role in inhibiting the fusion of phagosomes and lysosomes. Objective: the aim of this studyis to assess the difference of gene expression TACO protein in Latent Tuberculosis Infection (LTBI) and healthy people. Method: A preliminary studyof mRNA examination of TACO protein using Immunocytochemistry (ICC) and Real Time-Polimerase Chain Reaction (RT-PCR) method by a PCR Light Cycler 2.0 machine (Roche) in LTBI and healthy groups. Results: 18 samples of peripheral blood monocyte cells (PBMCs) were collected and divided into 2 groups. We found that there was a significantly difference between the 2 groups of samples. Conclusion: Further research is required to consider that the measurement of TACO expression using RT-PCRcan used as one of the other method to determine LTBI.

2016 ◽  
Vol 18 (2) ◽  
pp. 162
Author(s):  
Ryzky Widi Atmaja ◽  
Jusak Nugraha

Abstrak Latar Belakang. Imunitas memiliki peranan penting untuk melindungi host dari bacilli Mycobacterium tuberculosis (M.tb), bakteri Obligat  intraseluler  yang  menyebabkan  Tuberkulosis  (TB)  dan  latent  tuberculosis  infection  (LTBI).  Sel  T  subset  gamma-delta (T-γδ) adalah sel-sel potensial tersembunyi yang bermain peran di imunitas innate dan adaptive pada TB. Tetapi, hingga kini perananya   di   LTBI   masih   menjadi   misteri.   Bahan   dan   Metode.   Penelitian   dilakukan   dengan   melibatkan   10 penderita  TB serta 10 orang dengan LTBI. Mereka didapatkan dari Rumah Sakit Paru Surabaya melalui suatu persetujuan kelaikan etik   dari  Universitas  Airlangga.  Sampel-sampel  tersebut  akan  dihitung  jumlah  sel  T-γδ  menggunakan  F A C S C a l i b u r. Hasil.   Jumlah   sel   T-γδ   meningkat   pada   TB   (10,7%)   dan   LTBI   (15, 4%).   Jumlah   dari   kedua   kelompok   tersebut melebihi   rerata   normal   di   darah   tepi   (1% - 5%).   Kesimpulan.   Penigkatan   jumlah   sel   T-γδ   pada   TB   disebabkan melimpahnya kadar IL-12 yang dilepas oleh makrofag selama infeksi. Sementara, peningkatan jumlah sel T-γδ pada LTBI diasumsikan    karena    banyaknya    heat    shock    protein    (HSPs)    yang    dilepas    oleh    M.tb    di    bawah    kondisi    stres. ...Kata  kunci:  tuberkulosis,  latent  tuberculosis  infection,  Mycobacterium  tuberclosis,  sel  T  subset  gamma-d e l t a.


2018 ◽  
Vol 56 (11) ◽  
Author(s):  
Mi Ra Ryu ◽  
Min-Seung Park ◽  
Eun Hye Cho ◽  
Chul Won Jung ◽  
Kihyun Kim ◽  
...  

ABSTRACT QuantiFERON-TB Gold Plus (QFT-Plus) is a new-generation QuantiFERON-TB Gold In-Tube (QFT-GIT) assay which has two antigen-coated tubes called TB1, which contains long peptides derived from ESAT-6 and CFP-10, and TB2, which contains the same components as TB1 and additional short peptides which potentially stimulate CD8+ T cells through the presentation of major histocompatibility complex class I. This is the first study to compare QFT-Plus and QFT-GIT for use in the diagnosis of latent tuberculosis infection (LTBI) among immunocompromised patients in the Republic of Korea. Among 317 consecutive patients who underwent screening for LTBI before solid organ or hematopoietic stem cell transplantation and tumor necrosis factor alpha inhibitor treatment, LTBI was identified in 92 (29.0%) and 88 (27.8%) patients by QFT-GIT and QFT-Plus, respectively. The rate of concordance between QFT-GIT and QFT-Plus was 93.7% (κ value, 0.860), and the indeterminate rate (3.2%) was similar between QFT-GIT and QFT-Plus. Of 20 (6.3%) samples with discordant results, 11 (55.0%) and 7 (35.0%) were positive by QFT-GIT alone and QFT-Plus alone, respectively, and 2 (15.0%) were indeterminate by each assay. The interferon gamma level in samples with discordant results ranged from 0.39 to 1.10 IU/ml, except for one sample, in which the gamma interferon level was 2.97 IU/ml only in TB2. Conclusively, there was a high degree of agreement between the results of QFT-GIT and QFT-Plus for the screening of immunocompromised patients for LTBI. The reactivity in TB2 contributed substantially to the difference between QFT-GIT and QFT-Plus, particularly in solid organ transplant candidates. The significance of the discrete responses in TB1 and TB2 of QFT-Plus needs to be explored further by means of an immunological and clinical approach in different patient groups and clinical settings.


2015 ◽  
Vol 205 (1) ◽  
pp. 37-45 ◽  
Author(s):  
Ilaria Sauzullo ◽  
Fabio Mengoni ◽  
Claudia Mascia ◽  
Raffaella Rossi ◽  
Miriam Lichtner ◽  
...  

PLoS ONE ◽  
2013 ◽  
Vol 8 (3) ◽  
pp. e58378 ◽  
Author(s):  
Jimmy E. Rodríguez ◽  
Ana S. Ramírez ◽  
Laura P. Salas ◽  
Cecilia Helguera-Repetto ◽  
Jorge Gonzalez-y-Merchand ◽  
...  

Author(s):  
O.P. Lysenko ◽  
I.H. Vlasenko ◽  
V.V. Vlasenko ◽  
O.V. Rymsha ◽  
O.A. Nazarchuk ◽  
...  

Annotation. The relationship between tuberculosis infection and hemoblastosis has attracted the attention of physicians and has been the basis for a wide variety of assumptions. In the second half of the XIX century, there was expressed an opinion about the unique nature of these pathologies. They explained this phenomenon as the immunochemical affinity of specific tumor and mycobacterial antigens, but not as a result of the persistence of mycobacterium tuberculosis (MBT). The aim of the study was to investigate the association of latent tuberculosis infection and oncogenesis, in particular with leukemias. From transplantable human myeloblasts with acute myeloid leukemia (Kasumi-1) and T-lymphocytes with T-lymphoblastic leukemia (Jurkat), including 0.22 μm filtrate of their lysates were isolated from mycobacterium tuberculosis (MBT) with deficient cell wall (CWD). During primary microscopy, the MBT was found mainly in the form of coccoid. The isolated coccoid Kasumi and Jurkat had close antigen affinity and shared antigens with typical MBT. After prolonged incubation with the growth promoter, not only CWD MBT was isolated from all lysates and filtrates, but isolates with identical morphology and antigenic composition were obtained. This strongly suggested that the MBT and their modified forms could be associated with various types of leukemia. Moreover, the affinity of FLK-BLV isolates with blood of people with latent tuberculosis infection, cattle tuberculosis, CWD forms M. bovis and M. tuberculosis was determined, confirming the role of tuberculosis infection in the development of various forms of the disease depending on the host genome, as well as the features of the adaptive reactions of the infectious agent. In particular, after destruction by ultrasound and filtration through Amicon Ultracel® 100 K and Millex® GP 0.22 μm, their filtrates gave an increase in the number of CWD MBT with acid-resistant elements and had the same antigenic composition, including the original isolates. The long-term incubation of the test material in the growth promoter and numerous transplants on the adapted MyCel DW medium played a major role in the allocation of CWD.


2016 ◽  
Vol 6 (1) ◽  
pp. 12
Author(s):  
Rebekah Setiabudi ◽  
Ni Made Mertaniasih ◽  
Didik Didik Handijatmo ◽  
Retno Asih Setyoningrum

This study was intended to invent a simpler and more affordable method to establish diagnosis on Tuberculosis (TB) and Latent Tuberculosis infection (LTBI). Similar to “Quantiferon TB Gold In Tube” (QFT-GIT) and T.SPOT.TB methods, the researchers also utilized “early secreted antigenic target 6kDa” (ESAT-6) and “cultur filtrate protein 10kDa” (CFP-10) proteins to be induced on the specimen. ESAT-6 and CFP-10 are commercial products used to induce interferon gamma (INF-γ) which were to be read using sophisticated and expensive equipment. This study was intended to conduct an analysis on effective cocktail protein modification, i.e. ESAT-6, CFP-10 and Ag85A/B/C, with high validity to detect cellular immunity activity through in vitro examination on peripheral blood monocyte cells of Tuberculosis-suspected patients or patients with latent tuberculosis infection. Peripheral Blood Monocyte Cells (PBMCs) activity on children tuberculosis patient or Latent Tuberculosis Infection (LTBI), adult tuberculosis patient or LTBI, which induced by cocktail protein modification and not induced, were analyzed microscopically. The activity of PBMCs on children and adult tuberculosis patient or LTBI induced by RD1 secretory proteins: ESAT-6, CFP-10, Ag85A/B/C was higher compared to PBMCs which had not been induced by the secretory proteins. Cellular debris and monocyte cells with abnormal shapes were found on PBMCs which had been induced by RD1 secretory proteins at 8 th day after culture.


2019 ◽  
Vol 64 (5) ◽  
pp. 299-307
Author(s):  
M. V. Fursov ◽  
I. A. Dyatlov ◽  
V. D. Potapov

Modeling of tuberculosis infection is carried out in order to clarify various aspects of the tuberculosis pathogenesis, as well as the testing of new anti-tuberculosis drugs. The characteristic of in vitro models (n = 16) for Mycobacterium tuberculosis dormant state and in vivo models (n = 14) for the latent tuberculosis infection involving several animal species published to date are presented in this review. A brief description of the models and the results obtained by the authors are presented. The analysis of the published data reflects the list of methodological procedures that allow researchers to study the mechanism of the transition of M. tuberculosis cells to a dormant state and reverse to metabolically active state, as well as the process of conversion of active tuberculosis infection to a latent tuberculosis and reactivation.


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Ping-Huai Wang ◽  
Shu-Yung Lin ◽  
Susan Shih-Jung Lee ◽  
Shu-Wen Lin ◽  
Chih-Yuan Lee ◽  
...  

AbstractA significantly negative reversion in the QuantiFERON-TB Gold In-tube (QFT-GIT) test is reported in patients on dialysis, which makes the results unreliable. The CD4 and CD8 responses of the QFT-Gold plus (QFT-Plus) may have better positive consistency, but this needs to be investigated. We enrolled dialysis patients with baseline positive QFT-GIT0 results and conducted two rounds of follow-up paired QFT-GIT1&2 and QFT-Plus1&2 tests at an interval of 6 months. The positive consistency, concordance, and discordance of the QFT results were analyzed. A total of 236 patients on dialysis were screened, and 73 participants with positive QFT-GIT0 results were enrolled. The baseline QFT-GIT0 response was higher in the 1st QFT-Plus1(+) group than in the QFT-Plus1(−) group, but insignificantly different between the 1st QFT-GIT1(+) and QFT-GIT1(−) groups. The two assays had good correlation when concurrently tested. Fifty-three subjects completed a second round of the QFT-GIT2 and QFT-Plus2. Persistent positivity was higher with the QFT-Plus2 (81.8%) than with the QFT-GIT2 (58.8%, p = 0.040). The QFT-GIT1 and QFT-Plus1 CD4 responses were higher in patients with persistent positivity than in those with negative reversion, whereas the difference of the QFT-Plus TB1 and TB2 data, representative of the CD8 response, were similar between positive persistence and negative reversion. In conclusion, the QFT-Plus provides more reliable positive consistency than does the QFT-GIT. The CD4 interferon-γ response might play a role in maintaining positivity of LTBI.


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