scholarly journals Serologic Responses to Epitopes of the Major Surface Glycoprotein ofPneumocystis jiroveciDiffer in Human Immunodeficiency Virus–Infected and Uninfected Persons

2002 ◽  
Vol 186 (5) ◽  
pp. 644-651 ◽  
Author(s):  
Kieran R. Daly ◽  
Carl J. Fichtenbaum ◽  
Reiko Tanaka ◽  
Michael J. Linke ◽  
Robert O’Bert ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Surface Glycoprotein ◽  
Major Surface Glycoprotein ◽  
Immunodeficiency Virus ◽  
Major Surface

scholarly journals Analisis filogenetik gen mitochondrial large subunit (mtLSU) Pneumocystis jirovecii pada Pasien Human Immunodeficiency Virus (HIV) Terduga Pneumoniae di Jakarta

2020 ◽  
Vol 9 (1) ◽  
pp. 9-18
Author(s):  
Conny Riana Tjampakasari ◽  
Andi Yasmon ◽  
Agus Sjahrurachman ◽  
Samsuridjal Djauzi
Keyword(s):  
Human Immunodeficiency Virus ◽  
Hot Spot ◽  
Large Subunit ◽  
Surface Glycoprotein ◽  
Pneumocystis Jirovecii ◽  
Rt Pcr ◽  
Major Surface Glycoprotein ◽  
Immunodeficiency Virus ◽  
Polymerase Chain ◽  
Major Surface

Abstrak Pneumocystis jirovecii (P. jirovecii) merupakan patogen oportunistik yang penting pada pasien dengan gangguan kekebalan menurun khususnya human immunodeficiency virus (HIV). P. jirovecii tersebar dimanamana, menyebar melalui udara, dan menyerang sistem pernapasan atas. P. jirovecii mempunyai beberapa faktor virulensi, antara lain major surface glycoprotein (MSG) yang merupakan antigen yang paling banyak ditemukan di permukaan. Pendekatan biologi molekuler digunakan untuk mempelajari patogen ini karena hingga saat ini kultur belum dapat dilakukan. Penelitian ini bertujuan untuk memperoleh data genotip yang dapat dimanfaatkan sebagai dasar data demografi dan epidemiologi molekuler P. jirovecii di Indonesia. Dua puluh sampel sputum positif P. jirovecii pada real-time polymerase chain reaction (RT-PCR) dilakukan karakterisasi terhadap gen mitochondrial large subunit (mtLSU). Virulensi daerah hot spot gen mtLSU dianalisis dengan metode PCR dan sekuensing deoxyribonucleic acid (DNA). Diperoleh 30 strain dengan 7 varian didominasi oleh varian 3 yang bersirkulasi di Jakarta. Analisis filogenetik dengan strain negara lain menunjukkan strain Jakarta berkerabat dekat dengan strain Iran, India dan Korea. Kata kunci : Pneumocystis jirovecii, mtLSU, PCR, filogenetik Abstract Pneumocystis jirovecii (P. jirovecii) is an important opportunistic pathogen in immunocompromised patients especially human immunodeficiency virus (HIV). P. jirovecii is spread everywhere, spread through the air, and attacking the upper respiratory system. P. jirovecii has several virulence factors, including major surface glycoprotein (MSG) which is the most widely found on the surface antigen. The molecular biology approach is used to study this pathogen because until now culture cannot be done. This study aims to obtain genotype data that can be used as a basis for demographic and molecular epidemiological data of P. jirovecii in Indonesia. Twenty P. jirovecii positive sputum samples on real-time polymerase chain reaction (RT-PCR) were characterized by the mitochondrial large subunit (mtLSU) gene. Virulence of the mtLSU gene hot spot region was analyzed by PCR method and deoxyribonucleic acid (DNA) sequencing. Obtained 30 strains with 7 variants dominated by variant 3 circulating in Jakarta. Phylogenetic analyzed with strains of other countries shows that Jakarta strains are closely related to strains of Iran, India dan Korea. Keywords: Pneumocystis jirovecii, mtLSU, PCR, phylogenetic

scholarly journals Human Immunodeficiency Virus-Infected Patients with Prior Pneumocystis Pneumonia Exhibit Increased Serologic Reactivity to Several Major Surface Glycoprotein Clones

2006 ◽  
Vol 13 (10) ◽  
pp. 1071-1078 ◽  
Author(s):  
K. R. Daly ◽  
J. V. Koch ◽  
N. J. Shire ◽  
L. Levin ◽  
P. D. Walzer
Keyword(s):  
Human Immunodeficiency Virus ◽  
Blood Donors ◽  
The Other ◽  
Surface Glycoprotein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Immune Reactivity ◽  
Major Surface Glycoprotein ◽  
Immunodeficiency Virus ◽  
Major Surface ◽  
Hiv 1

ABSTRACT Recombinant clones of the carboxyl terminus of the major surface glycoprotein (MsgC) of Pneumocystis jirovecii are useful for analyzing serologic responses in humans. However, there is no standardized set of antigens in general use, which could lead to conflicting results. We have previously shown that human immunodeficiency virus type 1 (HIV-1)-infected patients with prior Pneumocystis pneumonia (PcP+) responded more frequently and more strongly to a clone of MsgC than did HIV-1-infected patients without PcP (PcP−). Here we test three new clones of MsgC to determine the effect of antigenic sequence variation on immune reactivity in blood donors and HIV-infected patients previously analyzed for reactivity to our original MsgC clone. In Western blot analyses, PcP+ patients exhibited the highest frequency of reactivity to each MsgC clone, and the frequency of reactivity with all four MsgC clones together was significantly higher in sera from PcP+ patients than in sera from the other patient groups. Furthermore, in an enzyme-linked immunosorbent assay we found that the PcP+ population had the highest level of reactivity to two of the four clones tested. One of the new clones could distinguish between PcP+ and PcP− populations, and two MsgC clones could distinguish blood donors from the other patient populations. The results show that inherent differences in MsgC amino acid sequence can affect recognition by antibodies independently of variations in protein length or patient population, and the utility of a clone depends on its sequence and on the populations tested.

scholarly journals Use of Oropharyngeal Washes to Diagnose and Genotype Pneumocystis jirovecii

2015 ◽  
Vol 2 (3) ◽  
Author(s):  
Jonathan J. Juliano ◽  
Eric Barnett ◽  
Christian M. Parobek ◽  
Steve M. Taylor ◽  
Steven R. Meshnick ◽  
...  
Keyword(s):  
Clinical Symptoms ◽  
High Sensitivity ◽  
Diagnostic Techniques ◽  
Surface Glycoprotein ◽  
Pneumocystis Jirovecii ◽  
Major Surface Glycoprotein ◽  
Immunodeficiency Virus ◽  
Noninvasive Samples ◽  
Strain Type ◽  
Major Surface

Abstract Pneumocystis jirovecii is a symbiotic respiratory fungus that presents in 2 clinical forms: pneumonia in immunocompromised patients or colonization, defined by the presence of the organism without associated clinical symptoms. Currently, diagnosis requires invasive bronchoscopy, which may not be available in some settings and is inappropriate for detecting colonization in healthy individuals. Noninvasive diagnostic techniques and molecular strain typing tools that can be used on these samples are critical for conducting studies to better understand transmission. We evaluated 2 real-time polymerase chain reaction (PCR) assays targeting dihydropteroate synthase and the major surface glycoprotein for detection in 77 oropharyngeal washes (OPWs) from 43 symptomatic human immunodeficiency virus-infected patients who underwent bronchoscopy. We also evaluated the ability of a new microsatellite (MS) genotyping panel to strain type infections from these samples. Each PCR used individually provided a high sensitivity (>80%) for detection of pneumonia but a modest specificity (<70%). When used in combination, specificity was increased to 100% with a drop in sensitivity (74%). Concentration of organisms by PCR in the OPW tended to be lower in colonized individuals compared with those with pneumonia, but differences in concentration could not clearly define colonization in symptomatic individuals. Oropharyngeal wash samples were genotyped using 6 MSs with ≥4 alleles successfully genotyped in the majority of colonized patients and ≥5 alleles in patients with pneumonia. The MS profile was consistent over time within patients with serial OPWs analyzed. Microsatellite genotyping on noninvasive samples may aid in studying the molecular epidemiology of this pathogen without requiring invasive diagnostic techniques.

scholarly journals Episomal Expression of Specific Sense and Antisense mRNAs in Leishmania amazonensis: Modulation of gp63 Level in Promastigotes and Their Infection of Macrophages In Vitro

2000 ◽  
Vol 68 (1) ◽  
pp. 80-86 ◽  
Author(s):  
De-Qiao Chen ◽  
Bala Krishna Kolli ◽  
Nagendra Yadava ◽  
Hong Gang Lu ◽  
Alice Gilman-Sachs ◽  
...  
Keyword(s):  
Kinetic Studies ◽  
Single Copy ◽  
Antisense Transcription ◽  
Leishmania Amazonensis ◽  
Surface Glycoprotein ◽  
Major Surface Glycoprotein ◽  
Wild Type Clone ◽  
Major Surface ◽  
Specific Sense

ABSTRACT The major surface glycoprotein (gp63) of Leishmania amazonensis is a metalloprotease implicated in the infection of mammalian macrophages. The expression of gp63 and its participation in this infection were further examined by modulating the level of this molecule in a virulent gp63-abundant wild-type clone. Promastigotes were transfected with gp63 genes cloned into aLeishmania-specific vector in two different orientations, leading to the expression of gp63 sense and antisense RNAs. With increasing selective pressure, cell surface gp63 was increasingly augmented in the transfectants with sense transcripts and suppressed to a very low level in those with antisense transcripts. Thus, the expression of gp63 from chromosomal, repetitive genes is not stringently regulated at the protein level and can be substantially reduced by episomal antisense transcription of a single copy. The transfectants differed significantly only in the level of gp63, thereby allowing specific evaluation of this molecule in leishmanial infection of macrophages in vitro. Kinetic studies of infection in vitro indicate that gp63 plays a role not only in the binding of this parasite to these macrophages but also in its intramacrophage survival and replication.

scholarly journals Amino-Terminal Substitutions in the CCR5 Coreceptor Impair gp120 Binding and Human Immunodeficiency Virus Type 1 Entry

1998 ◽  
Vol 72 (1) ◽  
pp. 279-285 ◽  
Author(s):  
Tatjana Dragic ◽  
Alexandra Trkola ◽  
Steven W. Lin ◽  
Kirsten A. Nagashima ◽  
Francis Kajumo ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Human Immunodeficiency Virus Type ◽  
Virus Type ◽  
Surface Glycoprotein ◽  
Alanine Scanning Mutagenesis ◽  
Cc Chemokine ◽  
Amino Terminal ◽  
Immunodeficiency Virus ◽  
Hiv 1

ABSTRACT The CC-chemokine receptor CCR5 is required for the efficient fusion of macrophage (M)-tropic human immunodeficiency virus type 1 (HIV-1) strains with the plasma membrane of CD4+ cells and interacts directly with the viral surface glycoprotein gp120. Although receptor chimera studies have provided useful information, the domains of CCR5 that function for HIV-1 entry, including the site of gp120 interaction, have not been unambiguously identified. Here, we use site-directed, alanine-scanning mutagenesis of CCR5 to show that substitutions of the negatively charged aspartic acid residues at positions 2 and 11 (D2A and D11A) and a glutamic acid residue at position 18 (E18A), individually or in combination, impair or abolish CCR5-mediated HIV-1 entry for the ADA and JR-FL M-tropic strains and the DH123 dual-tropic strain. These mutations also impair Env-mediated membrane fusion and the gp120-CCR5 interaction. Of these three residues, only D11 is necessary for CC-chemokine-mediated inhibition of HIV-1 entry, which is, however, also dependent on other extracellular CCR5 residues. Thus, the gp120 and CC-chemokine binding sites on CCR5 are only partially overlapping, and the former site requires negatively charged residues in the amino-terminal CCR5 domain.

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