In vivo tracking of th1 cells by PET reveals quantitative and temporal distribution and specific homing in lymphatic tissue

Christoph M. Griessinger, Rainer Kehlbach, Daniel Bukala, Stefan Wiehr, Rüdiger Bantleon, Funda Cay, Andreas Schmid, Heidi Braumüller, Birgit Fehrenbacher, Martin Schaller, Martin Eichner, Julie Sutcliffe, Walter Ehrlichmann, Oliver Eibl, Gerald Reischl, Simon R Cherry, Martin Röcken, Bernd J. Pichler, Manfred Kneilling

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Although T cells can be labeled for noninvasive in vivo imaging, little is known about the impact of such labeling on T-cell function, and most imaging methods do not provide holistic information about trafficking kinetics, homing sites, or quantification. Methods: We developed protocols that minimize the inhibitory effects of 64Cupyruvaldehyde- bis(N4- methylthiosemicarbazone) (64Cu-PTSM) labeling on T-cell function and permit the homing patterns of T cells to be followed by PET. Thus, we labeled ovalbumin (OVA) T-cell receptor transgenic interferon (IFN)-g-producing CD41 T (Th1) cells with 0.7-2.2 MBq of 64Cu-PTSM and analyzed cell viability, IFN-g production, proliferation, apoptosis, and DNA double-strand breaks and identified intracellular 64Cu accumulation sites by energy dispersive x-ray analysis. To elucidate the fate of Th1 cell homing by PET, 107 64Cu-OVA-Th1 cells were injected intraperitoneally or intravenously into healthy mice. To test the functional capacities of 64Cu-OVA-Th1 cells during experimental OVA-induced airway hyperreactivity, we injected 107 64Cu-OVA-Th1 cells intraperitoneally into OVA-immunized or nonimmunized healthy mice, which were challenged with OVA peptide or phosphate-buffered saline or remained untreated. In vivo PET investigations were followed by biodistribution, autoradiography, and fluorescence-activated cell sorting analysis. Results: PET revealed unexpected homing patterns depending on the mode of T-cell administration. Within 20 min after intraperitoneal administration, 64Cu-OVA-Th1 cells homed to the perithymic lymph nodes (LNs) of naive mice. Interestingly, intravenously administered 64Cu-OVA-Th1 cells homed predominantly into the lung and spleen but not into the perithymic LNs. The accumulation of 64Cu-OVA-Th1 cells in the pulmonary LNs (6.8 ± 1.1 percentage injected dose per cubic centimeter [%ID/cm3]) 24 h after injection was highest in the OVA-immunized and OVA-challenged OVA airway hyperreactivity-diseased littermates 24 h after intraperitoneal administration and lowest in the untreated littermates (3.7 ± 0.4 %ID/cm3). As expected, 64Cu-OVA-Th1 cells also accumulated significantly in the pulmonary LNs of nonimmunized OVA-challenged animals (6.1 ± 0.5 %ID/cm3) when compared with phosphate-buffered saline-challenged animals (4.6 ± 0.5%ID/cm3). Conclusion: Our protocol permits the detection of Th1 cells in single LNs and enables temporal in vivo monitoring of T-cell homing over 48 h. This work enables future applications for 64Cu-PTSM- labeled T cells in clinical trials and novel therapy concepts focusing on T-cell-based immunotherapies of autoimmune diseases or cancer.

Original languageEnglish (US)
Pages (from-to)301-307
Number of pages7
JournalJournal of Nuclear Medicine
Volume55
Issue number2
DOIs
StatePublished - Feb 1 2014

Fingerprint

Th1 Cells
Ovalbumin
Lymphoid Tissue
T-Lymphocytes
Lymph Nodes
Lung
Interferons
Phosphates
Double-Stranded DNA Breaks
T-Cell Antigen Receptor
Autoradiography
Immunotherapy

Keywords

  • In vivo cell tracking
  • Murine Th1 cells
  • Small animal PET

ASJC Scopus subject areas

  • Radiology Nuclear Medicine and imaging
  • Medicine(all)

Cite this

In vivo tracking of th1 cells by PET reveals quantitative and temporal distribution and specific homing in lymphatic tissue. / Griessinger, Christoph M.; Kehlbach, Rainer; Bukala, Daniel; Wiehr, Stefan; Bantleon, Rüdiger; Cay, Funda; Schmid, Andreas; Braumüller, Heidi; Fehrenbacher, Birgit; Schaller, Martin; Eichner, Martin; Sutcliffe, Julie; Ehrlichmann, Walter; Eibl, Oliver; Reischl, Gerald; Cherry, Simon R; Röcken, Martin; Pichler, Bernd J.; Kneilling, Manfred.

In: Journal of Nuclear Medicine, Vol. 55, No. 2, 01.02.2014, p. 301-307.

Research output: Contribution to journalArticle

Griessinger, CM, Kehlbach, R, Bukala, D, Wiehr, S, Bantleon, R, Cay, F, Schmid, A, Braumüller, H, Fehrenbacher, B, Schaller, M, Eichner, M, Sutcliffe, J, Ehrlichmann, W, Eibl, O, Reischl, G, Cherry, SR, Röcken, M, Pichler, BJ & Kneilling, M 2014, 'In vivo tracking of th1 cells by PET reveals quantitative and temporal distribution and specific homing in lymphatic tissue', Journal of Nuclear Medicine, vol. 55, no. 2, pp. 301-307. https://doi.org/10.2967/jnumed.113.126318
Griessinger, Christoph M. ; Kehlbach, Rainer ; Bukala, Daniel ; Wiehr, Stefan ; Bantleon, Rüdiger ; Cay, Funda ; Schmid, Andreas ; Braumüller, Heidi ; Fehrenbacher, Birgit ; Schaller, Martin ; Eichner, Martin ; Sutcliffe, Julie ; Ehrlichmann, Walter ; Eibl, Oliver ; Reischl, Gerald ; Cherry, Simon R ; Röcken, Martin ; Pichler, Bernd J. ; Kneilling, Manfred. / In vivo tracking of th1 cells by PET reveals quantitative and temporal distribution and specific homing in lymphatic tissue. In: Journal of Nuclear Medicine. 2014 ; Vol. 55, No. 2. pp. 301-307.
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abstract = "Although T cells can be labeled for noninvasive in vivo imaging, little is known about the impact of such labeling on T-cell function, and most imaging methods do not provide holistic information about trafficking kinetics, homing sites, or quantification. Methods: We developed protocols that minimize the inhibitory effects of 64Cupyruvaldehyde- bis(N4- methylthiosemicarbazone) (64Cu-PTSM) labeling on T-cell function and permit the homing patterns of T cells to be followed by PET. Thus, we labeled ovalbumin (OVA) T-cell receptor transgenic interferon (IFN)-g-producing CD41 T (Th1) cells with 0.7-2.2 MBq of 64Cu-PTSM and analyzed cell viability, IFN-g production, proliferation, apoptosis, and DNA double-strand breaks and identified intracellular 64Cu accumulation sites by energy dispersive x-ray analysis. To elucidate the fate of Th1 cell homing by PET, 107 64Cu-OVA-Th1 cells were injected intraperitoneally or intravenously into healthy mice. To test the functional capacities of 64Cu-OVA-Th1 cells during experimental OVA-induced airway hyperreactivity, we injected 107 64Cu-OVA-Th1 cells intraperitoneally into OVA-immunized or nonimmunized healthy mice, which were challenged with OVA peptide or phosphate-buffered saline or remained untreated. In vivo PET investigations were followed by biodistribution, autoradiography, and fluorescence-activated cell sorting analysis. Results: PET revealed unexpected homing patterns depending on the mode of T-cell administration. Within 20 min after intraperitoneal administration, 64Cu-OVA-Th1 cells homed to the perithymic lymph nodes (LNs) of naive mice. Interestingly, intravenously administered 64Cu-OVA-Th1 cells homed predominantly into the lung and spleen but not into the perithymic LNs. The accumulation of 64Cu-OVA-Th1 cells in the pulmonary LNs (6.8 ± 1.1 percentage injected dose per cubic centimeter [{\%}ID/cm3]) 24 h after injection was highest in the OVA-immunized and OVA-challenged OVA airway hyperreactivity-diseased littermates 24 h after intraperitoneal administration and lowest in the untreated littermates (3.7 ± 0.4 {\%}ID/cm3). As expected, 64Cu-OVA-Th1 cells also accumulated significantly in the pulmonary LNs of nonimmunized OVA-challenged animals (6.1 ± 0.5 {\%}ID/cm3) when compared with phosphate-buffered saline-challenged animals (4.6 ± 0.5{\%}ID/cm3). Conclusion: Our protocol permits the detection of Th1 cells in single LNs and enables temporal in vivo monitoring of T-cell homing over 48 h. This work enables future applications for 64Cu-PTSM- labeled T cells in clinical trials and novel therapy concepts focusing on T-cell-based immunotherapies of autoimmune diseases or cancer.",
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TY - JOUR

T1 - In vivo tracking of th1 cells by PET reveals quantitative and temporal distribution and specific homing in lymphatic tissue

AU - Griessinger, Christoph M.

AU - Kehlbach, Rainer

AU - Bukala, Daniel

AU - Wiehr, Stefan

AU - Bantleon, Rüdiger

AU - Cay, Funda

AU - Schmid, Andreas

AU - Braumüller, Heidi

AU - Fehrenbacher, Birgit

AU - Schaller, Martin

AU - Eichner, Martin

AU - Sutcliffe, Julie

AU - Ehrlichmann, Walter

AU - Eibl, Oliver

AU - Reischl, Gerald

AU - Cherry, Simon R

AU - Röcken, Martin

AU - Pichler, Bernd J.

AU - Kneilling, Manfred

PY - 2014/2/1

Y1 - 2014/2/1

N2 - Although T cells can be labeled for noninvasive in vivo imaging, little is known about the impact of such labeling on T-cell function, and most imaging methods do not provide holistic information about trafficking kinetics, homing sites, or quantification. Methods: We developed protocols that minimize the inhibitory effects of 64Cupyruvaldehyde- bis(N4- methylthiosemicarbazone) (64Cu-PTSM) labeling on T-cell function and permit the homing patterns of T cells to be followed by PET. Thus, we labeled ovalbumin (OVA) T-cell receptor transgenic interferon (IFN)-g-producing CD41 T (Th1) cells with 0.7-2.2 MBq of 64Cu-PTSM and analyzed cell viability, IFN-g production, proliferation, apoptosis, and DNA double-strand breaks and identified intracellular 64Cu accumulation sites by energy dispersive x-ray analysis. To elucidate the fate of Th1 cell homing by PET, 107 64Cu-OVA-Th1 cells were injected intraperitoneally or intravenously into healthy mice. To test the functional capacities of 64Cu-OVA-Th1 cells during experimental OVA-induced airway hyperreactivity, we injected 107 64Cu-OVA-Th1 cells intraperitoneally into OVA-immunized or nonimmunized healthy mice, which were challenged with OVA peptide or phosphate-buffered saline or remained untreated. In vivo PET investigations were followed by biodistribution, autoradiography, and fluorescence-activated cell sorting analysis. Results: PET revealed unexpected homing patterns depending on the mode of T-cell administration. Within 20 min after intraperitoneal administration, 64Cu-OVA-Th1 cells homed to the perithymic lymph nodes (LNs) of naive mice. Interestingly, intravenously administered 64Cu-OVA-Th1 cells homed predominantly into the lung and spleen but not into the perithymic LNs. The accumulation of 64Cu-OVA-Th1 cells in the pulmonary LNs (6.8 ± 1.1 percentage injected dose per cubic centimeter [%ID/cm3]) 24 h after injection was highest in the OVA-immunized and OVA-challenged OVA airway hyperreactivity-diseased littermates 24 h after intraperitoneal administration and lowest in the untreated littermates (3.7 ± 0.4 %ID/cm3). As expected, 64Cu-OVA-Th1 cells also accumulated significantly in the pulmonary LNs of nonimmunized OVA-challenged animals (6.1 ± 0.5 %ID/cm3) when compared with phosphate-buffered saline-challenged animals (4.6 ± 0.5%ID/cm3). Conclusion: Our protocol permits the detection of Th1 cells in single LNs and enables temporal in vivo monitoring of T-cell homing over 48 h. This work enables future applications for 64Cu-PTSM- labeled T cells in clinical trials and novel therapy concepts focusing on T-cell-based immunotherapies of autoimmune diseases or cancer.

AB - Although T cells can be labeled for noninvasive in vivo imaging, little is known about the impact of such labeling on T-cell function, and most imaging methods do not provide holistic information about trafficking kinetics, homing sites, or quantification. Methods: We developed protocols that minimize the inhibitory effects of 64Cupyruvaldehyde- bis(N4- methylthiosemicarbazone) (64Cu-PTSM) labeling on T-cell function and permit the homing patterns of T cells to be followed by PET. Thus, we labeled ovalbumin (OVA) T-cell receptor transgenic interferon (IFN)-g-producing CD41 T (Th1) cells with 0.7-2.2 MBq of 64Cu-PTSM and analyzed cell viability, IFN-g production, proliferation, apoptosis, and DNA double-strand breaks and identified intracellular 64Cu accumulation sites by energy dispersive x-ray analysis. To elucidate the fate of Th1 cell homing by PET, 107 64Cu-OVA-Th1 cells were injected intraperitoneally or intravenously into healthy mice. To test the functional capacities of 64Cu-OVA-Th1 cells during experimental OVA-induced airway hyperreactivity, we injected 107 64Cu-OVA-Th1 cells intraperitoneally into OVA-immunized or nonimmunized healthy mice, which were challenged with OVA peptide or phosphate-buffered saline or remained untreated. In vivo PET investigations were followed by biodistribution, autoradiography, and fluorescence-activated cell sorting analysis. Results: PET revealed unexpected homing patterns depending on the mode of T-cell administration. Within 20 min after intraperitoneal administration, 64Cu-OVA-Th1 cells homed to the perithymic lymph nodes (LNs) of naive mice. Interestingly, intravenously administered 64Cu-OVA-Th1 cells homed predominantly into the lung and spleen but not into the perithymic LNs. The accumulation of 64Cu-OVA-Th1 cells in the pulmonary LNs (6.8 ± 1.1 percentage injected dose per cubic centimeter [%ID/cm3]) 24 h after injection was highest in the OVA-immunized and OVA-challenged OVA airway hyperreactivity-diseased littermates 24 h after intraperitoneal administration and lowest in the untreated littermates (3.7 ± 0.4 %ID/cm3). As expected, 64Cu-OVA-Th1 cells also accumulated significantly in the pulmonary LNs of nonimmunized OVA-challenged animals (6.1 ± 0.5 %ID/cm3) when compared with phosphate-buffered saline-challenged animals (4.6 ± 0.5%ID/cm3). Conclusion: Our protocol permits the detection of Th1 cells in single LNs and enables temporal in vivo monitoring of T-cell homing over 48 h. This work enables future applications for 64Cu-PTSM- labeled T cells in clinical trials and novel therapy concepts focusing on T-cell-based immunotherapies of autoimmune diseases or cancer.

KW - In vivo cell tracking

KW - Murine Th1 cells

KW - Small animal PET

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