Development of a Multiplex Immunohistochemistry Workflow to Investigate the Immune Microenvironment in Mouse Models of Inflammatory Bowel Disease and Colon Cancer
被引:3
作者:
Pang, Lokman
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机构:
La Trobe Univ, Sch Canc Med, Olivia Newton John Canc Res Inst, Heidelberg, Vic 3084, AustraliaLa Trobe Univ, Sch Canc Med, Olivia Newton John Canc Res Inst, Heidelberg, Vic 3084, Australia
Pang, Lokman
[1
]
Ernst, Matthias
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La Trobe Univ, Sch Canc Med, Olivia Newton John Canc Res Inst, Heidelberg, Vic 3084, AustraliaLa Trobe Univ, Sch Canc Med, Olivia Newton John Canc Res Inst, Heidelberg, Vic 3084, Australia
Ernst, Matthias
[1
]
Huynh, Jennifer
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La Trobe Univ, Sch Canc Med, Olivia Newton John Canc Res Inst, Heidelberg, Vic 3084, AustraliaLa Trobe Univ, Sch Canc Med, Olivia Newton John Canc Res Inst, Heidelberg, Vic 3084, Australia
Huynh, Jennifer
[1
]
机构:
[1] La Trobe Univ, Sch Canc Med, Olivia Newton John Canc Res Inst, Heidelberg, Vic 3084, Australia
Multiplex immunohistochemistry (mIHC) enables simultaneous staining of multiple immune markers on a single tissue section. Mounting studies have demonstrated the versatility of mIHC in evaluating immune infiltrates in different diseases and the tumour microenvironment (TME). However, the majority of published studies are limited to the analysis of human patient samples. Performing mIHC on formalin-fixed paraffin-embedded (FFPE) mouse tissues, particularly with sensitive antigens, remain challenging. The aim of our study was to develop a robust and reproducible protocol to uncover the immune landscape in mouse FFPE tissues. Effective antibody stripping while maintaining sensitivity to antigens and tissue adhesion to the glass slide is critical in developing an mIHC panel to allow successive rounds of staining. Thus, we identified a highly efficient stripping method that preserves signal intensity and antigenicity to allow multiple rounds of staining. We subsequently optimised an mIHC workflow with antibodies specific against CD4, CD8 alpha, FOXP3 and B220 to identify distinct T and B cell populations on mouse FFPE tissues. Lastly, the application of this mIHC panel was validated in a mouse model of inflammatory bowel cancer, two allograft mouse models of spontaneous colon adenocarcinoma and a sporadic mouse model of colon cancer. Together, these demonstrate the utility of the aforementioned protocol in establishing the quantity and spatial localisation of immune cells in different pathological tissues.
机构:
Univ Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
Univ Calif Berkeley, Helen Wills Neurosci Inst, Berkeley, CA 94720 USA
Univ Calif Berkeley, Dept Integrat Biol, Berkeley, CA 94720 USAUniv Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
Ehrenberg, Alexander J.
;
Morales, Dulce Ovando
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Univ Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USAUniv Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
Morales, Dulce Ovando
;
Piergies, Antonia M. H.
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h-index: 0
机构:
Univ Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USAUniv Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
Piergies, Antonia M. H.
;
Li, Song Hua
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h-index: 0
机构:
Univ Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USAUniv Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
Li, Song Hua
;
Tejedor, Jorge Santos
论文数: 0引用数: 0
h-index: 0
机构:
Univ Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
Karolinska Inst, Dept Neurosci, Stockholm, SwedenUniv Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
Tejedor, Jorge Santos
;
Mladinov, Mihovil
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Univ Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USAUniv Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
Mladinov, Mihovil
;
Mulder, Jan
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机构:
Karolinska Inst, Dept Neurosci, Stockholm, SwedenUniv Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
Mulder, Jan
;
Grinberg, Lea T.
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机构:
Univ Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
Univ Sao Paulo, Sch Med, Sao Paulo, Brazil
Univ Calif San Francisco, Global Brain Hlth Inst, San Francisco, CA 94143 USAUniv Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
机构:
Nebraska Med Ctr, Dept Internal Med, Omaha, NE 68198 USANebraska Med Ctr, Dept Internal Med, Omaha, NE 68198 USA
Eichele, Derrick D.
;
Kharbanda, Kusum K.
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h-index: 0
机构:
Nebraska Med Ctr, Dept Internal Med, Omaha, NE 68198 USA
Western Iowa Hlth Care Syst, Vet Affairs Nebraska, Res Serv, Omaha, NE 68105 USA
Nebraska Med Ctr, Dept Biochem & Mol Biol, Omaha, NE 68198 USANebraska Med Ctr, Dept Internal Med, Omaha, NE 68198 USA
机构:
Providence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Oregon Hlth & Sci Univ, Dept Cell Dev & Canc Biol, Portland, OR 97239 USAProvidence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Feng, Zipei
;
Jensen, Shawn M.
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机构:
Providence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USAProvidence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Jensen, Shawn M.
;
Messenheimer, David J.
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h-index: 0
机构:
Providence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Oregon Hlth & Sci Univ, Dept Mol Microbiol & Immunol, Portland, OR 97239 USAProvidence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Messenheimer, David J.
;
Farhad, Mohammed
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h-index: 0
机构:
Providence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Oregon Hlth & Sci Univ, Dept Cell Dev & Canc Biol, Portland, OR 97239 USAProvidence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Farhad, Mohammed
;
Neuberger, Michael
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h-index: 0
机构:
Providence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Hosp Ludwig Maximilian Univ, Dept Gen Visceral Transplantat Vasc & Thorac Surg, Munich, GermanyProvidence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Neuberger, Michael
;
Bifulco, Carlo B.
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h-index: 0
机构:
Providence Portland Med Ctr, Dept Pathol, Portland, OR 97213 USAProvidence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Bifulco, Carlo B.
;
Fox, Bernard A.
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h-index: 0
机构:
Providence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Oregon Hlth & Sci Univ, Dept Mol Microbiol & Immunol, Portland, OR 97239 USAProvidence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
机构:
Univ Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
Univ Calif Berkeley, Helen Wills Neurosci Inst, Berkeley, CA 94720 USA
Univ Calif Berkeley, Dept Integrat Biol, Berkeley, CA 94720 USAUniv Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
Ehrenberg, Alexander J.
;
Morales, Dulce Ovando
论文数: 0引用数: 0
h-index: 0
机构:
Univ Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USAUniv Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
Morales, Dulce Ovando
;
Piergies, Antonia M. H.
论文数: 0引用数: 0
h-index: 0
机构:
Univ Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USAUniv Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
Piergies, Antonia M. H.
;
Li, Song Hua
论文数: 0引用数: 0
h-index: 0
机构:
Univ Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USAUniv Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
Li, Song Hua
;
Tejedor, Jorge Santos
论文数: 0引用数: 0
h-index: 0
机构:
Univ Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
Karolinska Inst, Dept Neurosci, Stockholm, SwedenUniv Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
Tejedor, Jorge Santos
;
Mladinov, Mihovil
论文数: 0引用数: 0
h-index: 0
机构:
Univ Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USAUniv Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
Mladinov, Mihovil
;
Mulder, Jan
论文数: 0引用数: 0
h-index: 0
机构:
Karolinska Inst, Dept Neurosci, Stockholm, SwedenUniv Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
Mulder, Jan
;
Grinberg, Lea T.
论文数: 0引用数: 0
h-index: 0
机构:
Univ Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
Univ Sao Paulo, Sch Med, Sao Paulo, Brazil
Univ Calif San Francisco, Global Brain Hlth Inst, San Francisco, CA 94143 USAUniv Calif San Francisco, Memory & Aging Ctr, Weill Inst Neuroscie Nces, San Francisco, CA 94143 USA
机构:
Nebraska Med Ctr, Dept Internal Med, Omaha, NE 68198 USANebraska Med Ctr, Dept Internal Med, Omaha, NE 68198 USA
Eichele, Derrick D.
;
Kharbanda, Kusum K.
论文数: 0引用数: 0
h-index: 0
机构:
Nebraska Med Ctr, Dept Internal Med, Omaha, NE 68198 USA
Western Iowa Hlth Care Syst, Vet Affairs Nebraska, Res Serv, Omaha, NE 68105 USA
Nebraska Med Ctr, Dept Biochem & Mol Biol, Omaha, NE 68198 USANebraska Med Ctr, Dept Internal Med, Omaha, NE 68198 USA
机构:
Providence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Oregon Hlth & Sci Univ, Dept Cell Dev & Canc Biol, Portland, OR 97239 USAProvidence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Feng, Zipei
;
Jensen, Shawn M.
论文数: 0引用数: 0
h-index: 0
机构:
Providence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USAProvidence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Jensen, Shawn M.
;
Messenheimer, David J.
论文数: 0引用数: 0
h-index: 0
机构:
Providence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Oregon Hlth & Sci Univ, Dept Mol Microbiol & Immunol, Portland, OR 97239 USAProvidence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Messenheimer, David J.
;
Farhad, Mohammed
论文数: 0引用数: 0
h-index: 0
机构:
Providence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Oregon Hlth & Sci Univ, Dept Cell Dev & Canc Biol, Portland, OR 97239 USAProvidence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Farhad, Mohammed
;
Neuberger, Michael
论文数: 0引用数: 0
h-index: 0
机构:
Providence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Hosp Ludwig Maximilian Univ, Dept Gen Visceral Transplantat Vasc & Thorac Surg, Munich, GermanyProvidence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Neuberger, Michael
;
Bifulco, Carlo B.
论文数: 0引用数: 0
h-index: 0
机构:
Providence Portland Med Ctr, Dept Pathol, Portland, OR 97213 USAProvidence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Bifulco, Carlo B.
;
Fox, Bernard A.
论文数: 0引用数: 0
h-index: 0
机构:
Providence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA
Oregon Hlth & Sci Univ, Dept Mol Microbiol & Immunol, Portland, OR 97239 USAProvidence Portland Med Ctr, Earle A Chiles Res Inst, Providence Canc Ctr, Robert W Franz Canc Res Ctr, Portland, OR 97213 USA