Microtiter enzyme-linked immunosorbent assay for immunoglobulin G cholera antitoxin in humans: method and correlation with rabbit skin vascular permeability factor technique

Young, Charles R.; Levine, Myron M.; Craig, James C.; Robins‐Browne, Roy M.

doi:10.1128/iai.27.2.492-496.1980

Cited by 32 publications

(19 citation statements)

References 18 publications

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“…Serology. Sera from volunteers who participated in the challenge were tested for IgG antibody to purified CS1 and purified CS3 by an enzyme-linked immunosorbent assay (67). These antigens (5 ,ug/ml) were applied to wells of polystyrene microtiter plates (Immulon I; Dynatech Laboratories, Inc., Alexandria, Va.).…”

mentioning

confidence: 99%

Coli surface antigens 1 and 3 of colonization factor antigen II-positive enterotoxigenic Escherichia coli: morphology, purification, and immune responses in humans

Levine¹,

Ristaino²,

Marley³

et al. 1984

Infect Immun

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Enterotoxigenic Escherichia coli (ETEC) of serotype 06:H16, biotype A, bearing colonization factor antigen II (CFA/II) possesses two distinct coli surface antigens, CS1 and CS3, whereas CFA/II-positive ETEC of serotype 08:H9 manifests only CS3. CS1 has been shown to be fimbrial in nature, but heretofore the morphology of CS3 has not been described. Accordingly, by immune electron microscopy we investigated the morphological characteristics of CS3 on bacterial cells and after purification. CS3 was found to consist of thin (2-nm), flexible, wiry, "fibrillar" fimbriae, visible both on bacteria (06:H16, biotype A, and 08:H9 strains) and in the pure state. In contrast, CS1 exists as wider (6-nm), rigid fimbriae on the surface of 06:H16, biotype A, strains. By the use of antisera to CS1 and CS3 in immune electron microscopy, immunodiffusion in gel, and immunoblotting techniques, CS1 and CS3 were found to be immunologically as well as morphologically distinct. Six of nine volunteers who developed diarrhea after challenge with an 0139:H28 ETEC strain bearing CS1 and CS3 had significant serological rises to purified CS1 and CS3 antigens, suggesting that both antigens are elaborated in vivo, play a role in pathogenesis, and stimulate an immune response.

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mentioning

confidence: 99%

Coli surface antigens 1 and 3 of colonization factor antigen II-positive enterotoxigenic Escherichia coli: morphology, purification, and immune responses in humans

Levine¹,

Ristaino²,

Marley³

et al. 1984

Infect Immun

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220

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“…Levels of vibriocidal antibody were measured against V. cholerae Ogawa strain 79 and Inaba strain 89 (6) by the microtiter technique, in which a fourfold or greater increase in titer is significant (1). Levels of IgG anti-cholera toxin were measured by an enzymelinked immunosorbent assay (ELISA) (11,17). Polystyrene plates with flat-bottomed wells were coated with 100 ,ul of 10-,ug/ml purified cholera toxin (Swiss Serum and Vaccine Institute, Berne, Switzerland) in Tris-EDTA buffer, pH 7.5.…”

Section: Methodsmentioning

confidence: 99%

“…IgA antibody to V. cholerae toxin and LPS (Inaba) was determined by ELISA. The assay was performed in the same manner as for sera except that the plates were washed seven times with phosphate-buffered saline-Tween (3 min per wash) (15,17).…”

Section: Methodsmentioning

confidence: 99%

Immunogenicity of two formulations of oral cholera vaccine comprised of killed whole vibrios and the B subunit of cholera toxin

et al. 1989

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Two formulations of oral cholera vaccine were evaluated for safety and immunogenicity in adult volunteers in Thailand, an area with sporadic cholera outbreaks. One formulation consisted of 2 x 1011 killed vibrios and 5 mg of cholera toxin B subunit, as was previously evaluated in North American volunteers, and the other consisted of 1 x 1011 killed vibrios and 1 mg of B subunit, as was recently evaluated in a field trial in Bangladesh. Three doses of each formulation were given with citrate-bicarbonate buffer. Neither formulation had adverse effects. The formulations stimulated similar serum immunoglobulin G (IgG) and IgA responses to Vibrio cholerae lipopolysaccharide and cholera toxin and intestinal secretory IgA responses to lipopolysaccharide and toxin after three doses. The formulation containing twice the quantity of killed vibrios stimulated better vibriocidal responses, especially to Ogawa serotype. A formulation of oral vaccine containing more killed vibrios than were included in the vaccine studied in the Bangladesh field trial may provide greater protection against cholera. Cholera provides considerable immunity against subsequent illness due to Vibrio cholerae for several years (7). However, parenteral vaccination with killed vibrio preparations offers only partial protection for less than 6 months (9, 13). With the recognition of the importance of local intestinal immunity against V. cholerae, attention has turned to the development of oral vaccines (9). In addition, with the understanding that cholera stimulates the production of both antibacterial and antitoxic antibodies in the intestine, emphasis has been given to combining elements in an oral vaccine to provoke an immune response against cholera toxin as well as against the organism itself (14, 15).Vaccines consisting of killed whole V. cholerae organisms and the purified B subunit of cholera toxin have been extensively evaluated. A single dose consisting of 2.5 mg of the B subunit and 5 x 1010 killed vibrios stimulated a local secretory immunoglobulin A (sIgA) antitoxin response and, in most, a sIgA antilipopolysaccharide (anti-LPS) response comparable to responses induced by cholera (15). Three doses, each comprising 5 mg of B subunit and 2 x 1011 killed vibrios or the killed vibrios alone, were studied for protective efficacy in North American adult volunteers (2). The combination oral vaccine provided 64% protection against cholera and the oral killed V. cholerae vaccine gave 56% protection. In a field trial subsequently performed in Bangladesh, an area where cholera is endemic, three doses of 1 mg of B subunit and 1011 killed whole vibrios were found to provide 62% efficacy, while the killed-vibrio vaccine alone gave 53% protection in the year after vaccination (3, 5). Since the formulation evaluated in volunteers and that tested in the field were never compared for immunogenicity, we chose to do so in adults in Thailand, an area with sporadic cholera outbreaks (recently due primarily to V. cholerae El Tor Inaba serotype). * Corresponding a...

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“…Sera were collected from all volunteers before the vaccination and on days 14, 26, 40, and 180. Levels of immunoglobulin G (IgG) cholera antitoxin were measured by an enzyme-linked immunosorbent assay (19).…”

Section: Methodsmentioning

confidence: 99%

Comparison of the reactivities and immunogenicities of procholeragenoid and the B subunit of cholera toxin in Thai volunteers

et al. 1989

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Procholeragenoid, a stable high-molecular-weight aggregate of cholera toxin derived by heat treatment, was evaluated for reactivity and immunogenicity in adult Thai volunteers. Since procholeragenoid is known to retain some residual activity of cholera toxin, increasing amounts were ingested until diarrhea occurred; 250 ,ug induced diarrhea, but 100 ,ug did not. Procholeragenoid and cholera toxin B subunit, both in 100-,ug amounts, were then compared for systemic and intestinal antitoxin responses. When three peroral doses were given, these immunogens gave comparable responses. The secretory immunoglobulin A antitoxin responses to three doses of 100 jig of B subunit did not differ significantly from responses found in previous studies of Thai adults given 1 or 5 mg of B subunit, but serum antitoxin responses were less after 1 or 2 doses of 100 jig than after doses of 1 or 5 mg. Serum antitoxin levels were similar after 3 doses of B subunit. Procholeragenoid in the maximum safe dose of 100 jig does not offer any immunologic advantage over B subunit, although it may be less expensive and easier to produce. However, these studies suggest that higher amounts of B subunit are more immunogenic and may be preferable, if found to be sufficiently cost effective, when added to oral killed whole Vibrio cholerae vaccines.

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Microtiter enzyme-linked immunosorbent assay for immunoglobulin G cholera antitoxin in humans: method and correlation with rabbit skin vascular permeability factor technique

Cited by 32 publications

References 18 publications

Coli surface antigens 1 and 3 of colonization factor antigen II-positive enterotoxigenic Escherichia coli: morphology, purification, and immune responses in humans

Coli surface antigens 1 and 3 of colonization factor antigen II-positive enterotoxigenic Escherichia coli: morphology, purification, and immune responses in humans

Immunogenicity of two formulations of oral cholera vaccine comprised of killed whole vibrios and the B subunit of cholera toxin

Comparison of the reactivities and immunogenicities of procholeragenoid and the B subunit of cholera toxin in Thai volunteers

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