Specific monoclonal antibodies and human immunoglobulin E show that Hev b 5 is an abundant allergen in high protein powdered latex gloves.
BACKGROUND
Hev b 5 is a major latex allergen recognized predominantly by latex-allergic health care workers (HCWs). Recombinant Hev b 5 (rHev b 5) was previously expressed as a fusion protein with maltose binding protein (MBP), itself an immunogenic molecule; therefore non-fusion rHev b 5 is desirable. Moreover, standardized immunological assays for the detection of Hev b 5 are currently lacking and may have important implications for both allergen avoidance and diagnosis in latex allergy.


OBJECTIVE
To generate and use Hev b 5-specific mAbs to determine the relative abundance of Hev b 5 in different latex extracts, correlating this with the IgE reactivity of latex-allergic HCWs and to produce non-fusion rHev b 5.


METHODS
For the production of mAbs, mice were immunized with rHev b 5/MBP fusion protein and mAbs selected with rHev b 5/MBP but not MBP reactivity. The mAb reactivity was compared with polyclonal IgE from latex-allergic HCWs using direct and inhibition ELISA and immunoblot assays. Recombinant Hev b 5 was expressed and purified in the pPROEX-HTa bacterial expression system.


RESULTS
Four Hev b 5-specific mAbs were produced. Immunoblotting and ELISA using the mAbs indicate abundant Hev b 5 in high protein powdered latex glove extracts as compared with crude latex sap extracts. High quality surgical gloves with no detectable protein have no detectable Hev b 5. Inhibition ELISAs using serum IgE from latex-allergic HCWs and Hev b 5-specific mAbs gave strong correlation. Non-fusion recombinant Hev b 5 was successfully expressed and purified, showing reactivity with both the Hev b 5-specific mAbs and serum IgE of latex-allergic HCWs.


CONCLUSIONS
Hev b 5-specific mAbs and human IgE from latex-allergic HCWs demonstrate the greater content of Hev b 5 in high protein powdered glove extracts. This may explain the observed higher frequency of sensitization to this allergen in HCWs.
Sutherland
M F
MF
Department of Allergy, Asthma and Clinical Immunology, The Alfred Hospital and Monash University, Victoria, Australia.
Drew
A
A
Rolland
J M
JM
Slater
J E
JE
Suphioglu
C
C
O'Hehir
R E
RE
D000485:Allergens / Q000032:analysis* / Q000235:genetics / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D052179:Antigens, Plant; D005932:Gloves, Surgical; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology*; D007840:Latex / Q000737:chemistry / Q000276:immunology; D020315:Latex Hypersensitivity / Q000276:immunology*; D010940:Plant Proteins; D011208:Powders; D011506:Proteins; D011994:Recombinant Proteins / Q000276:immunology
D000485:Allergens; D000911:Antibodies, Monoclonal; D052179:Antigens, Plant; D007840:Latex; D010940:Plant Proteins; D011208:Powders; D011506:Proteins; D011994:Recombinant Proteins; C101800:allergen Hev b 5; D007073:Immunoglobulin E