After blocking with 10% normal goat serum for 30 min at room temperature, these sections were incubated with rabbit anti-human CD66b
mAb (Abcam, dilution 1:200), rabbit anti-human MPO (Abcam, 1: 1000), or control IgG at 4[degrees]C overnight.
regulates adhesion and activation of human eosinophils," The Journal of Immunology, vol.
Sugihara et al., "Prognostic value of CD66b
positive tumor-infiltrating neutrophils in testicular germ cell tumor," BMC Cancer, vol.
Granulocytes were isolated from whole blood using a commercial kit for positive selection (StemCell Technologies Inc.), containing a tetrameric antibody complex that recognizes both the CD66b
antigen and the dextran of iron-coated nanoparticles.
Markers like CD62L low and CD11b high are associated with suppressor phenotypes, and others such as CD11c, CD32, CD35, CD45, and CD66b
can be upregulated in suppressor neutrophils.
Neutrophil infiltration was measured using a monoclonal-antibody stain against CD66b
, a granulocyte membrane antigen (about 96% of granulocytes in blood vessels are neutrophils).
Selectin "shedding" occurs because infected cells degranulate, including an EDTA-inhibitable sheddase (metalloprotease), [beta]2-integrins, CD66b
, and other inflammatory components such as matrix metalloproteases, which includes gelatinase (MMP9) (38,39).
(10.) Szekanecz Z, Haines GK, Harlow LA, et al: Increased synovial expression of the adhesion molecules CD66a, CD66b
and CD31 in rheumatoid and osteoarthritis.
Antibodies CEACAM1 mAb (283340), goat anti-mouse IgG (H + L), superclonal secondary antibody conjugated to Alexa Fluor 488, CD66b
mAb (G10F5) conjugated to FITC, and mouse IgM isotype control conjugated to FITC were from Invitrogen, Thermo Fisher Scientific.
This process involves chemokine receptors, such as CXCR2 (13), and the expression of receptors, such as CD11b and CD66b
, which are involved in the activation, migration, and adhesion of neutrophils (14).
We used 3-color analyses to identify granulocytes as sidescatter high, CD66b
high; monocytes as side-scatter intermediate, CD33 high; and T lymphocytes as sidescatter low, CD2 high.
Indeed, the expression of CD62L, a molecule of cell adhesion, is restored in patients receiving biological therapy, while expressions of CD11b (also called integrin alpha M) and CD66b
, another adhesion molecule, were decreased after treatment.