Allergy Asthma Immunol Res.  2015 Nov;7(6):583-589. 10.4168/aair.2015.7.6.583.

Blocking of Histamine Release and IgE Binding to FcepsilonRI on Human Basophils by Antibodies Produced in Camels

• Khaled AQ
• Sana Y
• Abdulrahman R
• Raida K
• Sami AH

• KMID: 2130265
• DOI: http://doi.org/10.4168/aair.2015.7.6.583

Abstract

PURPOSE
The production of camel heavy-chain antihuman IgE (huIgE) that has the potential to block IgE-FcepsilonRI interaction and histamine release by basophils.
METHODS
Camels were immunized with a synthetic loop peptide (SLP) designed in a multiple antigen peptide system (MAPS) forming SLP-MAPS immunogen. Camel polyclonal antibodies (PCAs) were produced, purified, characterized using Protein A & G, ELISA, and SDS-PAGE, and tested for their potency to block passive sensitization and histamine release of human basophils using flow cytometry (FCM) and ELISA, respectively.
RESULTS
FCM data indicated that camel conventional (IgG1) and heavy chain antibodies (HCAbs; IgG2, and IgG3) had blocking activities of 43.9%, 72%, and 96.6%, respectively. Moreover, both IgG2 and IgG3 achieved remarkable inhibition rates of 93.98% and 97.05% in histamine release, respectively, whereas the IgG1inhibiting activity was 60.05%.
CONCLUSIONS
Camel PCAs produced against SLP-MAPS were capable of blocking the IgE-receptor interaction and the release of histamine by basophils with superiority to HCAbs. These findings may pave the way toward the possible use of camel anti-huIgE HCAbs as blocking antibodies in the treatment of IgE-mediated allergy and asthma.

Keyword

Camels; antibodies; blocking; immunoglobulin E; anti-IgE antibodies; asthma; histamine release

MeSH Terms

Antibodies*
Antibodies, Blocking
Asthma
Basophils*
Camels*
Electrophoresis, Polyacrylamide Gel
Enzyme-Linked Immunosorbent Assay
Flow Cytometry
Histamine Release*
Histamine*
Humans*
Hypersensitivity
Immunoglobulin E*
Immunoglobulin G
Passive Cutaneous Anaphylaxis
Staphylococcal Protein A
Antibodies
Antibodies, Blocking
Histamine
Immunoglobulin E
Immunoglobulin G
Staphylococcal Protein A

Figure

• 

  Fig. 1 Kinetics of antibody response in camel immunized with SLP-MAPS. Camel was immunized 5 times at 2-week intervals with 0.5 mg/dose of SLP-MAPS in combination with Stimune adjuvant at 1:1 ratio.

• 

  Fig. 2 SDS-PAGE profile of purified camel anti-SLP-MAPS isotypes. Lanes: 1, IgG1; 2, IgG2; 3, IgG3; M, molecular weight markers.

• 

  Fig. 3 ELISA of binding activity of purified camel anti-SLP-MAPS isotypes. It shows the O.D of purified anti-SLP-MAPS compared to pre-and postimmunized sera.

• 

  Fig. 4 Flow cytometric analysis of the blocking potency of purified camel anti-SLP-MAPS isotypes to the binding of huIgE to its high affinity receptor (FcεRI) on stripped human basophils. A1-A4 & B1-B4. Fluorescence of anti-IgE-FITC to atopic serum-sensitized human basophils. In A1, atopic serum and camel isotypes are absent. B1 is a complete system in the absence of camel isotypes. B2-B4 Basophils sensitization in the presence of postimmunized camel IgG1 (B2), IgG2 (B3), and IgG3 (B4). A2-A4 Basophils sensitization in the presence of preimmunized camel IgG1 (A2), IgG2 (A3), and IgG3 (A4). Numbers in quadrants represent the percentages of IgE-positive basophils. Blot in the upper left corner of the histogram represent relevant dot blot.

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