Magnitude and kinetics of multifunctional CD4+ and CD8β+ T cells in pigs infected with swine influenza A virus

Talker, Stephanie C; Koinig, Hanna C; Stadler, Maria; Graage, Robert; Klingler, Eva; Ladinig, Andrea; Mair, Kerstin H; Hammer, Sabine E; Weissenböck, Herbert; Dürrwald, Ralf; Ritzmann, Mathias; Saalmüller, Armin; Gerner, Wilhelm

doi:10.1186/s13567-015-0182-3

Veterinary Research

Table 1 Antibody panels

From: Magnitude and kinetics of multifunctional CD4⁺ and CD8β⁺ T cells in pigs infected with swine influenza A virus

Antigen	Clone	Isotype	Fluorochrome	Labeling strategy	Source of primary Ab
Ex vivo Ki-67/CD4 ⁺ T cells
CD3	PPT3	IgG1	PE	secondary antibody^a	in house
CD4	74-12-4	IgG2b	Alexa488	secondary antibody^b	in house
CD8α	11/295/33	IgG2a	PE-Cy7	secondary antibody^c	in house
CD27	b30c7	IgG1	Alexa647	directly conjugated^d	in house
CD45RC	3a56	IgG1	PerCP-Cy5.5	directly conjugated^e	in house
SLA-DR	MSA3	IgG2a	Qdot605	directly conjugated	in house^f
Ki-67	B56	IgG1	V450	directly conjugated	BD Biosciences
Ex vivo Ki-67/CD8β ⁺ T cells
CD3	PPT3	IgG1	PE	secondary antibody^a	in house
CD8β	PG164A	IgG2a	Alexa488	Zenon labeling kit^g	VMRD
CD27	b30c7	IgG1	Alexa647	directly conjugated^d	in house
SLA-DR	MSA3	IgG2a	Qdot605	directly conjugated	in house^f
Perforin	δ-G9	IgG2b	PerCP-eFluor710	directly conjugated	eBioscience
Ki-67	B56	IgG1	V450	directly conjugated	BD Biosciences
Triple cytokine staining/CD4 ⁺ T cells
CD4	74-12-4	IgG2b	Alexa488	secondary antibody^b	in house
CD8α	11/295/33	IgG2a	PE-Cy7	secondary antibody^c	in house
CD27	b30c7	IgG1	BV421	biotin-streptavidin^h	in house
IFN-γ	P2G10	IgG1	PE	directly conjugated	BD Biosciences
TNF-α	MAb11	IgG1	BV605	directly conjugated	BioLegend
IL-2	A150D3F1	IgG2a	APC	directly conjugatedⁱ	Life Technologies
Triple cytokine staining/CD8β ⁺ T cells
CD3	BB23-8E6-8C8	IgG2a	PE-Cy7	directly conjugated	BD Biosciences
CD8β	PG164A	IgG2a	Alexa488	Zenon labeling kit^g	VMRD
CD27	b30c7	IgG1	BV421	biotin-streptavidin^h	in house
IFN-γ	P2G10	IgG1	PE	directly conjugated	BD Biosciences
TNF-α	MAb11	IgG1	BV605	directly conjugated	BioLegend
IL-2	A150D3F1	IgG2a	APC	directly conjugatedⁱ	Life Technologies
In vitro expanded CD8β ⁺ T cells
CD4	74-12-4	IgG2b	PerCP-Cy5.5	directly conjugated	BD Biosciences
CD8β	PG164A	IgG2a	Alexa488	secondary antibody^j	VMRD
CD107a	4E9/11	IgG1	Alexa647	directly conjugated	AbD Serotec
Pan-γδ	PPT16	IgG2b	PE-Cy7	biotin-streptavidin^k	in house
IFN-γ	P2G10	IgG1	PE	directly conjugated	BD Biosciences
TNF-α	MAb11	IgG1	BV605	directly conjugated	BioLegend

^aGoat anti-Mouse IgG1-PE, Southern Biotech.
^bGoat anti-Mouse IgG2b-Alexa488, Life Technologies.
^cGoat anti-Mouse IgG2a-PE-Cy7, Southern Biotech.
^dAlexa Fluor-647 Protein Labeling Kit, Life Technologies.
^eLightning-Link™ PerCP-Cy5.5 Tandem Conjugation Kit, Innova Biosciences.
^fCustom conjugation by Life Technologies.
^gIgG2a-Alexa488 Zenon labeling kit, Life Technologies.
^hStreptavidin-BV421, BioLegend.
ⁱLightning-Link™ APC Conjugation Kit, Innova Biosciences.
^jGoat anti-Mouse IgG2a-Alexa488, Life Technologies.
^kStreptavidin-PE-Cy7, eBioscience.

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ISSN: 1297-9716

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