Assessment of profilin as an allergen for latex-sensitized patients.
BACKGROUND
The presence of the actin-binding protein, profilin, has been demonstrated in natural latex extracts; but the clinical significance of this molecule as an allergen for latex-allergic patients is not clear. We studied the allergenic relevance of isolated latex natural and recombinant profilin, by in vivo and in vitro techniques, in two populations of spina bifida children (SB) and adults allergic to latex (AL).


METHODS
Profilin is present in small amounts in latex extracts obtained from low ammoniated (LA) natural latex. Its purification by affinity chromatography resulted difficult due to Heb v 1 unspecific binding. Therefore a method was developed to obtain natural profilin from natural latex, combining affinity chromatography (PLP, poly-L-proline Sepharose column) and previous ammonium sulfate fractionation. Alternatively, latex c-serum containing a low amount of Hev b 1 and a relatively higher profilin content could be used. Recombinant latex profilin isoform (rHev b 8) was cloned by PCR amplification. The entire coding region of Hev b 8 was subcloned into the expression vector pKN172 and a non-fusion form of Hev b 8 was expressed in Escherichia coli BL21 (DE3). Purified recombinant protein was obtained after a single passage through PLP-Sepharose column.


RESULTS
Natural and recombinant purified Hev b 8 were tested cutaneously by intradermoreaction (ID) in 17 SB and 14 AL patients. They were positive in 15 SB and 14 AL patients. No wheals were produced when tested in nonatopic control patients. Only 42% of sera from latex-allergic patients revealed specific IgE titers of class 1 or higher by enzyme immunoassay and only 39% of them exhibited IgE binding by SDS-PAGE immunoblotting with any natural or recombinant Hev b 8 forms.


CONCLUSIONS
It seems that profilin is a relevant allergen for both groups of patients from a frequency point of view, but with scarce presence in natural latex extracts and raw sources, with a subsequent low IgE induction capacity.
Nieto
A
A
Hospital Infantil La Fe, Valencia, Spain.
Mazón
A
A
Boquete
M
M
Carballada
F
F
Asturias
J A
JA
Martínez
J
J
Martínez
A
A
D000328:Adult; D000485:Allergens; D015153:Blotting, Western; D002648:Child; D003285:Contractile Proteins; D028482:Hevea / Q000737:chemistry; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood; D007840:Latex / Q000737:chemistry; D020315:Latex Hypersensitivity / Q000276:immunology*; D008840:Microfilament Proteins / Q000032:analysis / Q000276:immunology*; D010940:Plant Proteins / Q000276:immunology*; D051304:Profilins; D011852:Radioallergosorbent Test; D011994:Recombinant Proteins / Q000276:immunology; D012130:Respiratory Hypersensitivity / Q000276:immunology; D012882:Skin Tests; D016135:Spinal Dysraphism / Q000276:immunology
D000485:Allergens; D003285:Contractile Proteins; D007840:Latex; D008840:Microfilament Proteins; D010940:Plant Proteins; D051304:Profilins; D011994:Recombinant Proteins; D007073:Immunoglobulin E