Isolation of a Purified Allergen from Kentucky Blue Grass Pollen

Self Cite

An allergen, designated as Kentucky blue grass KBG-cytochrome c, was isolated from the nondialysable constituents (R) of the aqueous extract of KBG pollen by a combination of gel filtration and preparative isoelectrofocussing techniques. The allergen had an absorption spectrum characteristic of heme protein and had a molecular weight corresponding to 12,000 daltons. It was a basic protein with a pI value of 9.9 and contained all the common amino acids. KBG-cytochrome c elicited immediate hypersensitivity cutaneous reactions in individuals allergic to KBG pollen.

Section: Introductionmentioning

confidence: 83%

“…Preparation of R An aqueous extract of KBG pollen (HollisterStier Laboratory, Mississauga, Ont., Canada) was prepared according to the procedure previously established in this laboratory [7]. The aqueous ex tract was dialyzed through visking tubing, No.…”

Section: Methodsmentioning

confidence: 99%

Isolation of Allergenically Active Cytochrome c from Kentucky Blue Grass Pollen

Ekramoddoullah

Kisil

Sehon

1981

Self Cite

“…Murine antibodies to the crude extract of KBG pollen and [1-galactosidase were also produced by the same meth od. [20], The protein concentra tion of each extract was determined by Lowry's method [21].…”

Section: Preparation O F the Recombinant Allergen Rkbg83mentioning

confidence: 99%

Allergenic and Antigenic Cross-Reactivities of Group IX Grass Pollen Allergens

Zhang

Kisil²,

Sehon³

et al. 1991

The allergenic and antigenic cross-reactivities between a major recombinant Poa pratensis(Poa p) IX allergen, rKBG8.3, and its corresponding proteins of different grass pollens were examined. Immunoblotting of the proteins of thirteen different grass pollens using anti-rKBG8.3 antibodies indicated that Poa p IX-like proteins are present in ten other grass pollens, albeit in variable amounts and polymorphic forms. These proteins ranged in size from 20 to 88 kDa in different grass pollens. The percent relative binding determined for each grass pollen extract using allergic human sera showed a significant correlation (r = 0.891) with that of anti-rKBG8.3 antiserum. Moreover, there was a strong association (r = 0.901) between the Kentucky bluegrass extract and rKBG8.3 with respect to their inhibition of the binding of human IgE antibodies to allergens in grass pollen extracts. Taken together, these results suggest that the allergenic and antigenic epitopes of the Poa p IX-related proteins in some but not all grass pollens are similar in structure and specificities. It is concluded that the group IX allergens constitute a major family of homologous proteins in several grass pollens.

“…used in the initial phase of this study were pre pared as previously described [3,5]. However, the original method for the isolation of C-I-3 was sub stantially modified as outlined below.…”

Section: Preparation Of R and C-i-3mentioning

confidence: 99%

“…isolated by gel chromatography from the dialysate of KBG aq. ext., con tained components which were capable of suppressing both rat [3] and murine [4] IgE antibody responses to R, when D-I was ad ministered intraperitoneally prior to immu nization with R. Fraction D-I had been fur ther separated by ion-exchange chromato graphy and gel filtration into three fractions [5] and one of these, C-I-3, having an esti mated molecular weight of 4,800 daltons, was deemed to be devoid of determinants responsible for the allergenic activity of R, since it neither inhibited the binding of hu man anti-KBCi IgE antibodies to allcrgosorbent discs prepared with R nor triggered passive cutaneous anaphylaxis (PCA) in skin sites of hooded rats sensitized with mu rine or rat reaginic antibodies to R.…”

Section: Introductionmentioning

confidence: 99%

Isolation of a Low Molecular Weight Constituent from Kentucky Blue Grass Pollen Possessing Carrier Activity of High Molecular Weight Constituent(s)

Ekramoddoullah

Kisil

Sehon

1981

Self Cite

An immunologically active fraction, designated as C-I-3 was isolated from the dialysate of the aqueous extract of defatted Kentucky Blue Grass pollen by differential dialysis and gel filtration. This fraction was shown to consist of a glycoprotein with a molecular size corresponding to 4,800 daltons and with a pi value of 3.6, and to contain all amino acids except cysteine. The intraperitoneal administration of C-I-3 in saline into mice or rats prior to their immunization with the nondialyzable constituents of the aqueous extract of Kentucky Blue Grass (i.e., the retentate, R) in presence of Al(OH)₃ resulted in the enhanced production of anti-R IgE antibodies. The enhancement of the anti-R murine IgE response was shown in adoptive cell transfer experiments to be due to the generation of R-specific T helper cells. Hence, it is suggested that the structural elucidation of C-I-3 may prove useful in establishing the nature of carrier determinant(s) of the major allergenic components of Kentucky Blue Grass pollen.