cnr2 (cb2) ko Search Results


cb2 ko  (Jackson Laboratory)
86
Jackson Laboratory cb2 ko
WT and <t>CB2</t> KO mice were challenged intranasally for 3 days with rmIL-33. On day 4, lung ILC2s were FACS-sorted and cultured with rmIL-2 and rmIL-7 for 24 hours. (A-F) , RNA-seq was performed after 24 hours of culture. (A) Schematic overview of downstream CB2 regulatory signaling elements. (B) Heatmap illustrating gene set analysis (GSA) for RNA-seq expression, showing the differentially regulated genes in CB2 KO compared to WT ILC2s. Differentially expressed genes were selected based on p-value <0.05. (C) Ingenuity pathway analysis (IPA) results showing associated statistically significantly downregulated canonical pathways in CB2 KO ILC2s. Differentially regulated genes relevant to (D) ILC2 and CB2 signaling, (E) Cyclins and cell cycle regulation and (F) ILC2 proliferation signature. Differentially expressed genes were selected based on p-value <0.05. (G) ILC2 apoptosis and corresponding quantitation showing frequencies of early apoptotic (E.A., AnnexinV + DAPI − ) and late apoptotic/necrotic (L.A., AnnexinV + DAPI + ) ILC2s. Intranuclear expressions of (H) Bcl2, (I) Ki67, (J) pCREB, (K) pERK1/2, (L) p38, (M) pAKT and (N) GATA-3 and corresponding quantitation presented as MFI. Histograms show mean +/− SEM of a representative experiment of 2-3 with n=3-5. ns= non-significant, * p<0.05, ** p<0.01.
Cb2 Ko, supplied by Jackson Laboratory, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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balb cbyj  (Jackson Laboratory)
86
Jackson Laboratory balb cbyj
WT and <t>CB2</t> KO mice were challenged intranasally for 3 days with rmIL-33. On day 4, lung ILC2s were FACS-sorted and cultured with rmIL-2 and rmIL-7 for 24 hours. (A-F) , RNA-seq was performed after 24 hours of culture. (A) Schematic overview of downstream CB2 regulatory signaling elements. (B) Heatmap illustrating gene set analysis (GSA) for RNA-seq expression, showing the differentially regulated genes in CB2 KO compared to WT ILC2s. Differentially expressed genes were selected based on p-value <0.05. (C) Ingenuity pathway analysis (IPA) results showing associated statistically significantly downregulated canonical pathways in CB2 KO ILC2s. Differentially regulated genes relevant to (D) ILC2 and CB2 signaling, (E) Cyclins and cell cycle regulation and (F) ILC2 proliferation signature. Differentially expressed genes were selected based on p-value <0.05. (G) ILC2 apoptosis and corresponding quantitation showing frequencies of early apoptotic (E.A., AnnexinV + DAPI − ) and late apoptotic/necrotic (L.A., AnnexinV + DAPI + ) ILC2s. Intranuclear expressions of (H) Bcl2, (I) Ki67, (J) pCREB, (K) pERK1/2, (L) p38, (M) pAKT and (N) GATA-3 and corresponding quantitation presented as MFI. Histograms show mean +/− SEM of a representative experiment of 2-3 with n=3-5. ns= non-significant, * p<0.05, ** p<0.01.
Balb Cbyj, supplied by Jackson Laboratory, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/cnr2+(cb2)+ko/balb+cbyj/pmc09013728-45-1-23
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c57bl 6j  (Jackson Laboratory)
86
Jackson Laboratory c57bl 6j
WT and <t>CB2</t> KO mice were challenged intranasally for 3 days with rmIL-33. On day 4, lung ILC2s were FACS-sorted and cultured with rmIL-2 and rmIL-7 for 24 hours. (A-F) , RNA-seq was performed after 24 hours of culture. (A) Schematic overview of downstream CB2 regulatory signaling elements. (B) Heatmap illustrating gene set analysis (GSA) for RNA-seq expression, showing the differentially regulated genes in CB2 KO compared to WT ILC2s. Differentially expressed genes were selected based on p-value <0.05. (C) Ingenuity pathway analysis (IPA) results showing associated statistically significantly downregulated canonical pathways in CB2 KO ILC2s. Differentially regulated genes relevant to (D) ILC2 and CB2 signaling, (E) Cyclins and cell cycle regulation and (F) ILC2 proliferation signature. Differentially expressed genes were selected based on p-value <0.05. (G) ILC2 apoptosis and corresponding quantitation showing frequencies of early apoptotic (E.A., AnnexinV + DAPI − ) and late apoptotic/necrotic (L.A., AnnexinV + DAPI + ) ILC2s. Intranuclear expressions of (H) Bcl2, (I) Ki67, (J) pCREB, (K) pERK1/2, (L) p38, (M) pAKT and (N) GATA-3 and corresponding quantitation presented as MFI. Histograms show mean +/− SEM of a representative experiment of 2-3 with n=3-5. ns= non-significant, * p<0.05, ** p<0.01.
C57bl 6j, supplied by Jackson Laboratory, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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c b6 cg rag2 tm1 1 cgn j  (Jackson Laboratory)
86
Jackson Laboratory c b6 cg rag2 tm1 1 cgn j
WT and <t>CB2</t> KO mice were challenged intranasally for 3 days with rmIL-33. On day 4, lung ILC2s were FACS-sorted and cultured with rmIL-2 and rmIL-7 for 24 hours. (A-F) , RNA-seq was performed after 24 hours of culture. (A) Schematic overview of downstream CB2 regulatory signaling elements. (B) Heatmap illustrating gene set analysis (GSA) for RNA-seq expression, showing the differentially regulated genes in CB2 KO compared to WT ILC2s. Differentially expressed genes were selected based on p-value <0.05. (C) Ingenuity pathway analysis (IPA) results showing associated statistically significantly downregulated canonical pathways in CB2 KO ILC2s. Differentially regulated genes relevant to (D) ILC2 and CB2 signaling, (E) Cyclins and cell cycle regulation and (F) ILC2 proliferation signature. Differentially expressed genes were selected based on p-value <0.05. (G) ILC2 apoptosis and corresponding quantitation showing frequencies of early apoptotic (E.A., AnnexinV + DAPI − ) and late apoptotic/necrotic (L.A., AnnexinV + DAPI + ) ILC2s. Intranuclear expressions of (H) Bcl2, (I) Ki67, (J) pCREB, (K) pERK1/2, (L) p38, (M) pAKT and (N) GATA-3 and corresponding quantitation presented as MFI. Histograms show mean +/− SEM of a representative experiment of 2-3 with n=3-5. ns= non-significant, * p<0.05, ** p<0.01.
C B6 Cg Rag2 Tm1 1 Cgn J, supplied by Jackson Laboratory, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/cnr2+(cb2)+ko/1cgn+b6+cg+j+rag2+tm1/pmc09013728-45-4-23
Average 86 stars, based on 1 article reviews
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Image Search Results


WT and CB2 KO mice were challenged intranasally for 3 days with rmIL-33. On day 4, lung ILC2s were FACS-sorted and cultured with rmIL-2 and rmIL-7 for 24 hours. (A-F) , RNA-seq was performed after 24 hours of culture. (A) Schematic overview of downstream CB2 regulatory signaling elements. (B) Heatmap illustrating gene set analysis (GSA) for RNA-seq expression, showing the differentially regulated genes in CB2 KO compared to WT ILC2s. Differentially expressed genes were selected based on p-value <0.05. (C) Ingenuity pathway analysis (IPA) results showing associated statistically significantly downregulated canonical pathways in CB2 KO ILC2s. Differentially regulated genes relevant to (D) ILC2 and CB2 signaling, (E) Cyclins and cell cycle regulation and (F) ILC2 proliferation signature. Differentially expressed genes were selected based on p-value <0.05. (G) ILC2 apoptosis and corresponding quantitation showing frequencies of early apoptotic (E.A., AnnexinV + DAPI − ) and late apoptotic/necrotic (L.A., AnnexinV + DAPI + ) ILC2s. Intranuclear expressions of (H) Bcl2, (I) Ki67, (J) pCREB, (K) pERK1/2, (L) p38, (M) pAKT and (N) GATA-3 and corresponding quantitation presented as MFI. Histograms show mean +/− SEM of a representative experiment of 2-3 with n=3-5. ns= non-significant, * p<0.05, ** p<0.01.

Journal: The Journal of allergy and clinical immunology

Article Title: Cannabinoid Receptor II engagement promotes ILC2 expansion and enhances ILC2-dependent airway hyperreactivity

doi: 10.1016/j.jaci.2021.09.037

Figure Lengend Snippet: WT and CB2 KO mice were challenged intranasally for 3 days with rmIL-33. On day 4, lung ILC2s were FACS-sorted and cultured with rmIL-2 and rmIL-7 for 24 hours. (A-F) , RNA-seq was performed after 24 hours of culture. (A) Schematic overview of downstream CB2 regulatory signaling elements. (B) Heatmap illustrating gene set analysis (GSA) for RNA-seq expression, showing the differentially regulated genes in CB2 KO compared to WT ILC2s. Differentially expressed genes were selected based on p-value <0.05. (C) Ingenuity pathway analysis (IPA) results showing associated statistically significantly downregulated canonical pathways in CB2 KO ILC2s. Differentially regulated genes relevant to (D) ILC2 and CB2 signaling, (E) Cyclins and cell cycle regulation and (F) ILC2 proliferation signature. Differentially expressed genes were selected based on p-value <0.05. (G) ILC2 apoptosis and corresponding quantitation showing frequencies of early apoptotic (E.A., AnnexinV + DAPI − ) and late apoptotic/necrotic (L.A., AnnexinV + DAPI + ) ILC2s. Intranuclear expressions of (H) Bcl2, (I) Ki67, (J) pCREB, (K) pERK1/2, (L) p38, (M) pAKT and (N) GATA-3 and corresponding quantitation presented as MFI. Histograms show mean +/− SEM of a representative experiment of 2-3 with n=3-5. ns= non-significant, * p<0.05, ** p<0.01.

Article Snippet: C57BL/6J, BALB/cByJ, Rag2 −/− (C.B6(Cg)-Rag2 tm1.1Cgn /J), Rag2 −/− IL-2Rg −/− (C;129S4-Rag2tm1.1Flv Il2rgtm1.1Flv/J) and CB2 KO (B6.129P2- Cnr2 tm1Dgen /J) were purchased from The Jackson Laboratory and bred in our animal facility at the Keck School of Medicine, University of Southern California (USC).

Techniques: Cell Culture, RNA Sequencing, Expressing, Quantitation Assay

(A) Lung ILC2s were gated as single live CD45 + , Lineage − , ST2 + and CD127 + cells, expressing high levels of GATA-3. (B-H) C57BL/6 (WT) or CB2 KO mice were challenged intranasally for 3 days with rmIL-33 or PBS. On day 4, lung ILC2s were isolated by FACS. (B) CB2 expression on ILC2s and (C) corresponding quantitation presented as CB2 MFI. On day 4, analysis of lung function, BAL cellular content and lung histology were performed. (D) Lung resistance and (E) dynamic compliance in response to methacholine. (F) Total BAL CD45 + cells and (G) BAL cellularity. (H) Lung H&E staining. (I-O) WT or CB2 KO mice were challenged intranasally for 5 days with A. alternata or PBS. On day 6, lung ILC2s were isolated by FACS. (I) CB2 expression on ILC2s and (J) corresponding quantitation presented as CB2 MFT On day 6, analysis of lung function, BAL cellular content and lung histology were performed. (K) Lung resistance and (L) dynamic compliance in response to methacholine. (M) Total BAL CD45 + cells and (N) BAL cellularity. (O) Lung H&E staining. Histograms show mean +/− SEM of a representative experiment of 3 with n=4-9. Scale bars 50μm. eosinophils (eos.), polymorphonuclear neutrophils (PMN). ns= non-significant, * p<0.05.

Journal: The Journal of allergy and clinical immunology

Article Title: Cannabinoid Receptor II engagement promotes ILC2 expansion and enhances ILC2-dependent airway hyperreactivity

doi: 10.1016/j.jaci.2021.09.037

Figure Lengend Snippet: (A) Lung ILC2s were gated as single live CD45 + , Lineage − , ST2 + and CD127 + cells, expressing high levels of GATA-3. (B-H) C57BL/6 (WT) or CB2 KO mice were challenged intranasally for 3 days with rmIL-33 or PBS. On day 4, lung ILC2s were isolated by FACS. (B) CB2 expression on ILC2s and (C) corresponding quantitation presented as CB2 MFI. On day 4, analysis of lung function, BAL cellular content and lung histology were performed. (D) Lung resistance and (E) dynamic compliance in response to methacholine. (F) Total BAL CD45 + cells and (G) BAL cellularity. (H) Lung H&E staining. (I-O) WT or CB2 KO mice were challenged intranasally for 5 days with A. alternata or PBS. On day 6, lung ILC2s were isolated by FACS. (I) CB2 expression on ILC2s and (J) corresponding quantitation presented as CB2 MFT On day 6, analysis of lung function, BAL cellular content and lung histology were performed. (K) Lung resistance and (L) dynamic compliance in response to methacholine. (M) Total BAL CD45 + cells and (N) BAL cellularity. (O) Lung H&E staining. Histograms show mean +/− SEM of a representative experiment of 3 with n=4-9. Scale bars 50μm. eosinophils (eos.), polymorphonuclear neutrophils (PMN). ns= non-significant, * p<0.05.

Article Snippet: C57BL/6J, BALB/cByJ, Rag2 −/− (C.B6(Cg)-Rag2 tm1.1Cgn /J), Rag2 −/− IL-2Rg −/− (C;129S4-Rag2tm1.1Flv Il2rgtm1.1Flv/J) and CB2 KO (B6.129P2- Cnr2 tm1Dgen /J) were purchased from The Jackson Laboratory and bred in our animal facility at the Keck School of Medicine, University of Southern California (USC).

Techniques: Expressing, Isolation, Quantitation Assay, Staining

(A) BM ILC2 progenitors gated as CLPs (CD45 + Lin − CD127 + α4β7 − Flt3 + ), αLPs (CD45 + Lin − CD127 + α4β7 + Flt3 − ), ChILPs (CD45 + Lin CD127 + α4β7 + Flt3 − CD25 − ), ILC2Ps (CD45 + LiN − CD127 + α4β7 + Flt3 − CD25 + ). (B) Frequencies within CD45 + BM cells of CLPs, αLPs, ChILPs and ILC2Ps in naïve WT and CB2 KO mice. (C) Numbers of ST2 + ILC2s in the BM (one femur). (D-L) WT or CB2 KO mice were challenged intranasally for 3 days with rmIL-33 or PBS and on day 4, lung ILC2 numbers, activation, apoptosis and proliferation were analyzed. (D) Live CD45 + Lin − CD127 + ST2 + lung ILC2s and (E) corresponding quantitation showing the numbers of ILC2s per lung. (F) Early apoptotic (E.A., AnnexinV + DAPT − ) and late apoptotic/necrotic (L.A., AnnexinV + DAPI + ) ILC2s (black dots: WT mice, red dots: CB2 KO mice) and (G) corresponding quantitation showing frequencies of E.A. and L.A. (H) Lung ILC2 intranuclear Ki67 expression and (I) corresponding quantitation showing Ki67 MFI. (J) IL-13 (top panel) and IL-5 (bottom panel) positive ILC2s and corresponding quantitation of (K) IL-13- and (L) IL-5-positive lung ILC2s, showing frequencies of protein expression. Histograms show mean +/− SEM of a representative experiment of 3 with n=4-14. ns= non-significant, * p<0.05. FMO: Full-minus-one.

Journal: The Journal of allergy and clinical immunology

Article Title: Cannabinoid Receptor II engagement promotes ILC2 expansion and enhances ILC2-dependent airway hyperreactivity

doi: 10.1016/j.jaci.2021.09.037

Figure Lengend Snippet: (A) BM ILC2 progenitors gated as CLPs (CD45 + Lin − CD127 + α4β7 − Flt3 + ), αLPs (CD45 + Lin − CD127 + α4β7 + Flt3 − ), ChILPs (CD45 + Lin CD127 + α4β7 + Flt3 − CD25 − ), ILC2Ps (CD45 + LiN − CD127 + α4β7 + Flt3 − CD25 + ). (B) Frequencies within CD45 + BM cells of CLPs, αLPs, ChILPs and ILC2Ps in naïve WT and CB2 KO mice. (C) Numbers of ST2 + ILC2s in the BM (one femur). (D-L) WT or CB2 KO mice were challenged intranasally for 3 days with rmIL-33 or PBS and on day 4, lung ILC2 numbers, activation, apoptosis and proliferation were analyzed. (D) Live CD45 + Lin − CD127 + ST2 + lung ILC2s and (E) corresponding quantitation showing the numbers of ILC2s per lung. (F) Early apoptotic (E.A., AnnexinV + DAPT − ) and late apoptotic/necrotic (L.A., AnnexinV + DAPI + ) ILC2s (black dots: WT mice, red dots: CB2 KO mice) and (G) corresponding quantitation showing frequencies of E.A. and L.A. (H) Lung ILC2 intranuclear Ki67 expression and (I) corresponding quantitation showing Ki67 MFI. (J) IL-13 (top panel) and IL-5 (bottom panel) positive ILC2s and corresponding quantitation of (K) IL-13- and (L) IL-5-positive lung ILC2s, showing frequencies of protein expression. Histograms show mean +/− SEM of a representative experiment of 3 with n=4-14. ns= non-significant, * p<0.05. FMO: Full-minus-one.

Article Snippet: C57BL/6J, BALB/cByJ, Rag2 −/− (C.B6(Cg)-Rag2 tm1.1Cgn /J), Rag2 −/− IL-2Rg −/− (C;129S4-Rag2tm1.1Flv Il2rgtm1.1Flv/J) and CB2 KO (B6.129P2- Cnr2 tm1Dgen /J) were purchased from The Jackson Laboratory and bred in our animal facility at the Keck School of Medicine, University of Southern California (USC).

Techniques: Activation Assay, Quantitation Assay, Expressing

(A-E) FACS-sorted WT and CB2 KO activated ILC2s (aILC2) were adoptively transferred to Rag2 −/− IL-2Rg −/− mice (5x10 4 ILC2s/mouse), and then challenged intranasally for 3 days with rmIL-33 or PBS. On day 4, lung function, BAL eosinophilia and lung ILC2 numbers were analyzed. (A) Experimental design. (B) Lung resistance and (C) dynamic compliance in response to methacholine. (D) BAL eosinophils and (E) lung ILC2s. (F-M) WT (CD45.1) and CB2 KO (CD45.2) mice were challenged intranasally for 3 days with rmIL-33. On day 4, lung ILC2s were FACS-sorted and 5x10 4 of a 1:1 mix of CD45.1:CD45.2 ILC2s/mouse was adoptively transferred to Rag2 −/− IL-2Rg −/− mice. Host mice were then challenged intranasally for 3 days with rmIL-33 and lung ILC2 numbers, survival and proliferation were measured. (F) Experimental design. (G) Ratio of transferred CD45.1:CD45.2 aILC2s. (H) Total lung ILC2s 24h and 72h after transfer and (I) corresponding quantitation showing ILC2 numbers per lung. (J) CD45.1 and CD45.2 ILC2 fold-increase over 48h (24h to 72h post transfer). (K) ILC2 Ki67 MFI and (L) frequencies of live CD45.1 and CD45.2 ILC2s 24h and 72h post transfer. Histograms show mean +/− SEM of a representative experiment of 2 with n=5. ns= non-significant, * p<0.05, *** p<0.001. NT: no transfer.

Journal: The Journal of allergy and clinical immunology

Article Title: Cannabinoid Receptor II engagement promotes ILC2 expansion and enhances ILC2-dependent airway hyperreactivity

doi: 10.1016/j.jaci.2021.09.037

Figure Lengend Snippet: (A-E) FACS-sorted WT and CB2 KO activated ILC2s (aILC2) were adoptively transferred to Rag2 −/− IL-2Rg −/− mice (5x10 4 ILC2s/mouse), and then challenged intranasally for 3 days with rmIL-33 or PBS. On day 4, lung function, BAL eosinophilia and lung ILC2 numbers were analyzed. (A) Experimental design. (B) Lung resistance and (C) dynamic compliance in response to methacholine. (D) BAL eosinophils and (E) lung ILC2s. (F-M) WT (CD45.1) and CB2 KO (CD45.2) mice were challenged intranasally for 3 days with rmIL-33. On day 4, lung ILC2s were FACS-sorted and 5x10 4 of a 1:1 mix of CD45.1:CD45.2 ILC2s/mouse was adoptively transferred to Rag2 −/− IL-2Rg −/− mice. Host mice were then challenged intranasally for 3 days with rmIL-33 and lung ILC2 numbers, survival and proliferation were measured. (F) Experimental design. (G) Ratio of transferred CD45.1:CD45.2 aILC2s. (H) Total lung ILC2s 24h and 72h after transfer and (I) corresponding quantitation showing ILC2 numbers per lung. (J) CD45.1 and CD45.2 ILC2 fold-increase over 48h (24h to 72h post transfer). (K) ILC2 Ki67 MFI and (L) frequencies of live CD45.1 and CD45.2 ILC2s 24h and 72h post transfer. Histograms show mean +/− SEM of a representative experiment of 2 with n=5. ns= non-significant, * p<0.05, *** p<0.001. NT: no transfer.

Article Snippet: C57BL/6J, BALB/cByJ, Rag2 −/− (C.B6(Cg)-Rag2 tm1.1Cgn /J), Rag2 −/− IL-2Rg −/− (C;129S4-Rag2tm1.1Flv Il2rgtm1.1Flv/J) and CB2 KO (B6.129P2- Cnr2 tm1Dgen /J) were purchased from The Jackson Laboratory and bred in our animal facility at the Keck School of Medicine, University of Southern California (USC).

Techniques: Quantitation Assay

(A-C) BALB/c mice were challenged intranasally for 3 days with rmIL-33 or PBS. On day 4, lung ILC2s were FACS-sorted and cultured with rmIL-2 and rmIL-7 with or without 10μg/mL CB2 agonist for 24 hours. (A) Levels of IL-5, IL-13 in the culture supernatants. (B) ILC2 apoptosis and corresponding quantitation showing frequencies of early apoptotic (E.A., AnnexinV + DAPI − ) and late apoptotic/necrotic (L.A., AnnexinV + DAPI + ) ILC2s. (C) ILC2 Ki67 expression and corresponding quantitation showing Ki67 MFI. (D-O) Rag2 −/− mice were challenged with rmIL-33 for 3 days intranasally with CB2 agonist (JWH133, 1mg/kg) or vehicle and euthanized on day 4. (D) Experimental design. (E) Lung resistance in response to methacholine. (F) Lung H&E staining, scale bars 50μm. (G) Total BAL CD45 + cells and (H) BAL eosinophils. (I) Live lung ILC2s and (J) corresponding quantitation showing the numbers of ILC2s per lung. (K) E.A. and L.A. ILC2s and corresponding quantitation showing frequencies of E.A. and L.A. (L) Lung ILC2 Ki67 expression and corresponding quantitation showing Ki67 MFI. (M) IL-5 and (O) IL-13 positive ILC2s and corresponding quantitation (N, P) , showing frequencies of protein expression. Histograms show mean +/− SEM of a representative experiment of 3 with n=4-6. ns= non-significant, * p<0.05, ** p<0.01.

Journal: The Journal of allergy and clinical immunology

Article Title: Cannabinoid Receptor II engagement promotes ILC2 expansion and enhances ILC2-dependent airway hyperreactivity

doi: 10.1016/j.jaci.2021.09.037

Figure Lengend Snippet: (A-C) BALB/c mice were challenged intranasally for 3 days with rmIL-33 or PBS. On day 4, lung ILC2s were FACS-sorted and cultured with rmIL-2 and rmIL-7 with or without 10μg/mL CB2 agonist for 24 hours. (A) Levels of IL-5, IL-13 in the culture supernatants. (B) ILC2 apoptosis and corresponding quantitation showing frequencies of early apoptotic (E.A., AnnexinV + DAPI − ) and late apoptotic/necrotic (L.A., AnnexinV + DAPI + ) ILC2s. (C) ILC2 Ki67 expression and corresponding quantitation showing Ki67 MFI. (D-O) Rag2 −/− mice were challenged with rmIL-33 for 3 days intranasally with CB2 agonist (JWH133, 1mg/kg) or vehicle and euthanized on day 4. (D) Experimental design. (E) Lung resistance in response to methacholine. (F) Lung H&E staining, scale bars 50μm. (G) Total BAL CD45 + cells and (H) BAL eosinophils. (I) Live lung ILC2s and (J) corresponding quantitation showing the numbers of ILC2s per lung. (K) E.A. and L.A. ILC2s and corresponding quantitation showing frequencies of E.A. and L.A. (L) Lung ILC2 Ki67 expression and corresponding quantitation showing Ki67 MFI. (M) IL-5 and (O) IL-13 positive ILC2s and corresponding quantitation (N, P) , showing frequencies of protein expression. Histograms show mean +/− SEM of a representative experiment of 3 with n=4-6. ns= non-significant, * p<0.05, ** p<0.01.

Article Snippet: C57BL/6J, BALB/cByJ, Rag2 −/− (C.B6(Cg)-Rag2 tm1.1Cgn /J), Rag2 −/− IL-2Rg −/− (C;129S4-Rag2tm1.1Flv Il2rgtm1.1Flv/J) and CB2 KO (B6.129P2- Cnr2 tm1Dgen /J) were purchased from The Jackson Laboratory and bred in our animal facility at the Keck School of Medicine, University of Southern California (USC).

Techniques: Cell Culture, Quantitation Assay, Expressing, Staining

(A) Human blood ILC2s (hILC2s) were FACS-sorted from PBMCs and cultured for 72h with rhIL-2 and rhIL-7 with or without rhIL-33. (B) Flow cytometry plots of isolated hILC2s. (C) CB2 expression on hILC2s cultured with or without rhIL-33 and (D) corresponding quantitation showing CB2 MFI. (E-H) Isolated hILC2s were cultured for 72h with rhIL-2, rhIL-7, rhIL-33 with or without (E) CB2 antagonist (AM630, 10μg/mL) or (G) CB2 agonist (JWH133, 10μg/mL). Levels of IL-5, IL-13 and IL-6 in the culture supernatants following (F) CB2 antagonist and (H) CB2 agonist treatment. Histograms show 4 independent healthy blood donors. ns= non-significant, * p<0.05, ** p<0.01. (I) Isolated hILC2s were cultured for 72h with rhIL-2, rhIL-7, and rhIL-33 prior to adoptive transfer of 50,000 hILC2s in Rag2 −/− IL-2Rg −/− mice. Host mice were then challenged intranasally for 3 days with rhIL-33 and either CB2 antagonist or agonist (both 1mg/kg) or the vehicle (control). On day 4, lung function, BAL eosinophils and lung hILC2s were measured. (J) Lung resistance in response to methacholine. Number of (K) BAL eosinophils and (L) hILC2s per lung. Histograms show mean +/− SEM, n=4. ns= non-significant, * p<0.05, ** p<0.01. Data representative of 2 independent experiments.

Journal: The Journal of allergy and clinical immunology

Article Title: Cannabinoid Receptor II engagement promotes ILC2 expansion and enhances ILC2-dependent airway hyperreactivity

doi: 10.1016/j.jaci.2021.09.037

Figure Lengend Snippet: (A) Human blood ILC2s (hILC2s) were FACS-sorted from PBMCs and cultured for 72h with rhIL-2 and rhIL-7 with or without rhIL-33. (B) Flow cytometry plots of isolated hILC2s. (C) CB2 expression on hILC2s cultured with or without rhIL-33 and (D) corresponding quantitation showing CB2 MFI. (E-H) Isolated hILC2s were cultured for 72h with rhIL-2, rhIL-7, rhIL-33 with or without (E) CB2 antagonist (AM630, 10μg/mL) or (G) CB2 agonist (JWH133, 10μg/mL). Levels of IL-5, IL-13 and IL-6 in the culture supernatants following (F) CB2 antagonist and (H) CB2 agonist treatment. Histograms show 4 independent healthy blood donors. ns= non-significant, * p<0.05, ** p<0.01. (I) Isolated hILC2s were cultured for 72h with rhIL-2, rhIL-7, and rhIL-33 prior to adoptive transfer of 50,000 hILC2s in Rag2 −/− IL-2Rg −/− mice. Host mice were then challenged intranasally for 3 days with rhIL-33 and either CB2 antagonist or agonist (both 1mg/kg) or the vehicle (control). On day 4, lung function, BAL eosinophils and lung hILC2s were measured. (J) Lung resistance in response to methacholine. Number of (K) BAL eosinophils and (L) hILC2s per lung. Histograms show mean +/− SEM, n=4. ns= non-significant, * p<0.05, ** p<0.01. Data representative of 2 independent experiments.

Article Snippet: C57BL/6J, BALB/cByJ, Rag2 −/− (C.B6(Cg)-Rag2 tm1.1Cgn /J), Rag2 −/− IL-2Rg −/− (C;129S4-Rag2tm1.1Flv Il2rgtm1.1Flv/J) and CB2 KO (B6.129P2- Cnr2 tm1Dgen /J) were purchased from The Jackson Laboratory and bred in our animal facility at the Keck School of Medicine, University of Southern California (USC).

Techniques: Cell Culture, Flow Cytometry, Isolation, Expressing, Quantitation Assay, Adoptive Transfer Assay, Control