Characterization of antigen-presenting cells in human apical periodontitis lesions by flow cytometry and immunocytochemistry.
OBJECTIVE
To analyse phenotypic characteristics of antigen-presenting cells (APC), isolated from human periapical lesions by flow cytometry and immunocytochemistry.


METHODS
Sixteen periapical lesions were digested for 15 min with 0.05% collagenase. Mononuclear cells, separated from other inflammatory cells by density centrifugation, were processed for flow cytometry and/or immunocytochemistry. Single and double immunostainings were performed using monoclonal antibodies specific for human CD45, CD3, CD19, CD14, HLA-DR, CD1a, CD83 and CD123.


RESULTS
Antigen-presenting cells (HLA-DR(+) cells) represented 32.9 +/- 17.8% of total mononuclear cells. Amongst them, B cells (HLA-DR(+) CD19(+)) were the predominant APC population, followed by activated macrophages (HLA-DR(+) CD14(+)), dendritic cells (DC) (HLA-DR(+) CD14(-) CD19(-) CD3(-)) and activated T cells (HLA-DR(+) CD3(+)). Based on the predominance of T cells (CD3(+)) or B cells and plasma cells (CD19(+) and CD19(lo), respectively) amongst mononuclear cell infiltrates, lesions were divided into T- and B-types. The percentage of DC in T-type lesions (27.1 +/- 6.8% of total HLA-DR(+) cells) was higher, compared with B-type lesions (10.3 +/- 5.2%) (P < 0.01). Within the DC population, the percentages of CD1a (Langerhans cell type) and CD123 (probably plasmacytoid DC type) did not differ significantly between the groups (P > 0.05). However, the percentage of mature DC (CD83(+)) was significantly higher in T-type periapical lesions (P < 0.05).


CONCLUSIONS
Flow cytometry and immunocytochemistry are suitable methods for phenotypic analysis of APC after their isolation from human periapical lesions. APC, that were phenotypically heterogeneous, constituted a significant component of infiltrating cells. Lesions with the predominance of T cells were characterized by a higher proportion of mature DC (HLA-DR(+)CD83(+) cells) than lesions with predominance of B cells/plasma cells.
Lukić
A
A
Department of Endodontics, Faculty of Stomatology, University of Belgrade, Belgrade, Serbia and Montenegro.
Vasilijić
S
S
Majstorović
I
I
Vucević
D
D
Mojsilović
S
S
Gazivoda
D
D
Danilović
V
V
Petrović
R
R
Colić
M
M
D000293:Adolescent; D000328:Adult; D000938:Antigen-Presenting Cells / Q000145:classification / Q000473:pathology*; D015703:Antigens, CD / Q000032:analysis; D018949:Antigens, CD1 / Q000032:analysis; D018941:Antigens, CD19 / Q000032:analysis; D001402:B-Lymphocytes / Q000473:pathology; D017252:CD3 Complex / Q000032:analysis; D003713:Dendritic Cells / Q000473:pathology; D005434:Flow Cytometry; D006684:HLA-DR Antigens / Q000032:analysis; D006801:Humans; D007136:Immunoglobulins / Q000032:analysis; D007150:Immunohistochemistry; D016130:Immunophenotyping; D053650:Interleukin-3 Receptor alpha Subunit; D017493:Leukocyte Common Antigens / Q000032:analysis; D018950:Lipopolysaccharide Receptors / Q000032:analysis; D008213:Lymphocyte Activation; D008264:Macrophages / Q000473:pathology; D008562:Membrane Glycoproteins / Q000032:analysis; D008875:Middle Aged; D010485:Periapical Periodontitis / Q000276:immunology / Q000473:pathology*; D010950:Plasma Cells / Q000473:pathology; D016185:Receptors, Interleukin-3 / Q000032:analysis; D013601:T-Lymphocytes / Q000473:pathology; D000097802:CD83 Antigen
D015703:Antigens, CD; D018949:Antigens, CD1; D018941:Antigens, CD19; D017252:CD3 Complex; D006684:HLA-DR Antigens; D007136:Immunoglobulins; D053650:Interleukin-3 Receptor alpha Subunit; D017493:Leukocyte Common Antigens; D018950:Lipopolysaccharide Receptors; D008562:Membrane Glycoproteins; D016185:Receptors, Interleukin-3; D000097802:CD83 Antigen; C419393:CD1a antigen; C115022:IL3RA protein, human