Radioimmunoprecipitation and Western Blotting with Sera of Human Immunodeficiency Virus Infected Patients: A Comparative Study

Chiodi, Francesca; Bredberg-Rådén, Ulla; Biberfeld, Gunnel; Böttiger, Blenda; Albert, Jan; Åsjö, Birgitta; Fenyõ, Éva Mária; Norrby, Erling

doi:10.1089/aid.1987.3.165

Cited by 38 publications

(14 citation statements)

References 23 publications

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“…Certainly, a replicating virus is a more powerful inducer for MIF release than a soluble glycoprotein, through a mechanism not yet described. Taking in account that the gp120 circulates in AIDS patients (Chiodi et al, 1987), this viral protein may contribute to increasing systemic MIF levels as we describe here. Since MIF secretion can be induced by a variety of endogenous/exogenous stimuli (Calandra and Roger, 2003), we understand that it is very difficult to determine the real nature of MIF release in vivo .…”

Section: Discussionmentioning

confidence: 95%

Elevated levels of macrophage migration inhibitory factor (MIF) in the plasma of HIV-1-infected patients and in HIV-1-infected cell cultures: A relevant role on viral replication

et al. 2010

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The cytokine macrophage migration inhibitory factor (MIF) is involved in the pathogenesis of inflammatory and infectious diseases, however its role in HIV-1 infection is unknown. Here we show that HIV-1-infected patients present elevated plasma levels of MIF, that HIV-1-infected peripheral blood mononuclear cells (PBMCs) release a greater amount of MIF, and that the HIV-1 envelope glycoprotein gp120 induces MIF secretion from uninfected PBMCs. The HIV-1 replication in PBMCs declines when these cells were treated with anti-MIF antibodies, or when treated with the ABC-transporter inhibitor probenecid, which also inhibited MIF secretion. The addition of recombinant MIF (rhMIF) to HIV-1-infected PBMCs enhances viral replication of CCR5- or CXCR4-tropic HIV-1 isolates. Using a T CD4+ cell lineage containing an HIV long terminal repeats (LTR)-Luciferase construct, we detected that rhMIF promotes transcription from HIV-1 LTR. Our results show that HIV-1 induces MIF secretion and suggest that MIF influences the HIV-1 biology through activation of HIV-1 LTR.

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Section: Discussionmentioning

confidence: 95%

Elevated levels of macrophage migration inhibitory factor (MIF) in the plasma of HIV-1-infected patients and in HIV-1-infected cell cultures: A relevant role on viral replication

et al. 2010

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“…McKendali et al (1988) Blumberg et al (1987);Chiodi et al (1987); Knuver et al(1987);Thorpe et al (1987);Blomberg and Klasse (1988);Grimaldi et al (1988);Heberling et al (1988);Tersmette et al (1988) Ulstrup et al (1986;Yamaguchi et al et al (1985);Fang et al (1986);Ohta et al (1986) (…”

mentioning

confidence: 99%

Immunoblotting and dot blotting

Stott

1989

Journal of Immunological Methods

166

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“…Reactivity to core proteins only as been reported to represent a source of non-specificity in the WB test (3.). Nevertheless, RIPA has previously been shown to ensure a high degree of specificity in HIV serology (8), and false positive results are uncommon by this method. The unconstant reactivity could be explained by sample deterioration during long storage period; alternatively, high IgG concentration (4) may be the source of non-specifiticy.…”

Section: Resultsmentioning

confidence: 99%

“…492 nm values equal or greater than 0.2 in triplicate tests were considered as positive. Positive specimens and a randomly selected group of negative specimens were assayed by radioimmunoprecipitation assay (RIPA) and Western blot (WB; Bio-Rad Clinical Division) test as previously described (8). Antigen for the H1V-1 confirmatory tests was obtained from HTLV-IIIB infected cells; HTLV-IV and SBL-6669 strains were used as antigens (1) in the HIV-2 RIPA and WB test.…”

Section: Methodsmentioning

confidence: 99%

Screening of African sera stored for more than 17 years for HIV antibodies by site-directed serology

Chiodi

Biberfeld²,

Parks

et al. 1989

Eur J Epidemiol

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Antibodies to Human Immunodeficiency Virus type 1 (HIV-1) and type 2 (HIV-2) were investigated, using site-directed enzyme immunoassay (ELISA), in 320 specimens obtained from three remote, African tribes during 1969-1971. Using HIV-1 E34/E32 ELISA and HIV-2 149 ELISA, assay were conducted on 101 serum specimens from the Korekore tribe of Zimbabwe, 93 specimens from the Mano tribe of Liberia, and 126 specimens from the Turkana tribe of Kenya; specimens which tested positive in ELISA were further tested by radioimmunoprecipitation assay (RIPA) and Western blot (WB). Two serum specimens from the Mano tribe of Liberia gave OD 492 nm values greater than 0.2 in HIV E34/E32 ELISA in all three runs. These two specimens reacted with HIV-1 envelope proteins gp160 and gp120 and the internal protein p24 in RIPA and WB; however, the reactivity was uncostant. All other serum specimens were negative for HIV-1 and HIV-2 antibodies. Site directed ELISA serology for HIV-1 and HIV-2 gave very low rates of false positive reactivity. Thus, reaction with HIV-1 antigen was identified in two persons of one tribe in Liberia in 1971, but HIV-2 antibodies were not detected in this tribe; HIV-1 and HIV-2 antibodies were absent during the late 1960's and early 1970's from two African tribes resident in Zimbabwe and Kenya.

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Radioimmunoprecipitation and Western Blotting with Sera of Human Immunodeficiency Virus Infected Patients: A Comparative Study

Cited by 38 publications

References 23 publications

Elevated levels of macrophage migration inhibitory factor (MIF) in the plasma of HIV-1-infected patients and in HIV-1-infected cell cultures: A relevant role on viral replication

Elevated levels of macrophage migration inhibitory factor (MIF) in the plasma of HIV-1-infected patients and in HIV-1-infected cell cultures: A relevant role on viral replication

Immunoblotting and dot blotting

Screening of African sera stored for more than 17 years for HIV antibodies by site-directed serology

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