Stereological quantification of carboxyfluorescein-labeled rat lung metastasis: a new method for the assessment of natural killer cell activity and tumor adhesion in vivo and in situ

Hörsten, Stephan von; Helfritz, Andreas; Kuhlmann, Susanne; Nave, Heike; Tschernig, Thomas; Pabst, Reinhard; Ben‐Eliyahu, Shamgar; Meyer, Dirk; Schmidt, Reinhold; Schmitz, Christoph

doi:10.1016/s0022-1759(00)00162-9

Cited by 26 publications

(20 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…16 Previous research points to the importance of NKCC in clearing MADB106 cells from the lung. Specifically, pulmonary NK cells interact with intravenously inoculated MADB106 cells in situ; 30 treatments that enhance NKCC (e.g., lipopolysacchardie or Poly I-C administration) improve tumor clearance and reduce metastasis; 23 manipulations that compromise NKCC (e.g., hypothermia, alcohol consumption, food restriction and tobacco smoke components) interfere with resistance to MADB106 metastasis; [31][32][33][34] and selective depletion of NK cells typically increases the lung reten- tion of MADB106 more than 100-fold. 27,32,35 Thus, although factors other than NKCC, like B-cell and macrophage activity, 36,37 may modulate lung MADB106 metastasis, it appears that lysis of MADB106 cells by NK cells is a major factor mediating shortterm clearance of this tumor from the lungs.…”

Section: Discussionmentioning

confidence: 99%

“…This population of cells is critical for recognizing and destroying MADB106 cells that have infiltrated the lungs. 30,34 If, as some suggest, a tradeoff in cell numbers exists between the blood on the one hand and the spleen and lungs on the other, 17 then one might expect the number of pulmonary NK cells to exhibit circadian changes similar to those affecting splenic NK cells.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Diurnal changes in lung tumor clearance and their relation to NK cell cytotoxicity in the blood and spleen

2001

View full text Add to dashboard Cite

Natural killer cell cytotoxicity (NKCC) was reported to manifest a circadian rhythm, peaking during wakefulness in both human blood and rat spleen. Using F344 rats, we investigated whether such fluctuations (i) reflect changes in NK cell numbers or in cytotoxicity per cell; (ii) coincide in the blood and spleen; (iii) correspond with clearance of NK-sensitive tumor cells from the lungs and (iv) influence formation of lung metastases. Two rat groups were housed in opposite 12:12 hr lighting regimens. Two hours after the onset of light or dark, both groups were either sacrificed or intravenously inoculated with tumor cells to study the following indices: NKCC and NK cell numbers in the spleen (n ‫؍‬ 29) and blood (n ‫؍‬ 79), lung clearance of tumor cells (n ‫؍‬ 142) and lung metastasis (n ‫؍‬ 69). The tumor employed, MADB106, is an NK-sensitive mammary adenocarcinoma that metastasizes only to the lungs. The results indicated that, during the dark phase, splenic NKCC increased (37% higher lytic unit [LU] 50 ) mostly due to a 28.9% higher percentage of NK cells in the spleen. In contrast, blood NKCC decreased by 42.5% (LU 20 ) and this decline was independent of circulating NK cell numbers, which remained constant. Lung tumor clearance increased in the dark (up to 42% lower retention 9 hr after inoculation), but no corresponding changes in the number of metastases were observed 3 weeks later. We conclude that diurnal changes in rats' NKCC are organ-specific, involve changes in both cell distribution and activity and may affect short-term in vivo indices of NK tumoricidal activity. Natural killer (NK) cells, discovered 25 years ago, 1 are a subset of lymphocytes that spontaneously recognize and lyse virally infected and malignant cells. 2 In vivo, they are believed to play a substantial role in controlling the metastasis of several malignancies. This premise is based on the better prognosis associated with higher NK cell cytotoxicity (NKCC) in cancer patients 3 and is corroborated by several animal models that demonstrate a critical role for NK cells in controlling experimentally induced metastasis. 4,5 A number of immunologic indices, such as the production of several cytokines and the numbers and activity of various immune cells, show circadian fluctuations of appreciable magnitude, 6,7 but the in vivo ramifications of these fluctuations are largely unknown. A circadian rhythm of NKCC in humans has been documented by a substantial number of studies, most of which recorded increased NKCC in the blood during the day. 8 -11 Most of these studies used the standard NKCC assay, which measures NKCC per a fixed number of peripheral blood mononuclear cells (PBMCs). Because the percentage of NK cells within PBMCs also peaks during the day, 10,12,13 it is not clear how much of the rise in NKCC results from an increased proportion of NK cells within PBMCs and how much from changes in the cytotoxicity of individual NK cells.In rats, NKCC has been studied in the spleen. 14 -16 These studies located the maximal and minimal level...

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Diurnal changes in lung tumor clearance and their relation to NK cell cytotoxicity in the blood and spleen

2001

View full text Add to dashboard Cite

show abstract

“…Recent studies demonstrate that stress hormones such as adrenaline modulate the lung retention of MADB106 tumor cells even within minutes. 7 Thus, these data suggest that detailed time kinetic studies of the cellular changes within the first minutes and hours will provide new insights into specific mechanism determining the initial clearance rate of tumor cells in target organs and for the interaction of different leukocyte subsets in the early surveillance of metastasis.…”

mentioning

confidence: 94%

“…The present approach combines immunohistochemic methods with vital dye labeling techniques and takes advantage of the visualization and quantification of effector-like cells that co-localized with CFSE (5-(and -6)-carboxyfluorescein diacetate succinimidyl ester) positive MADB106 tumor targets in lung tissue in situ. 7 Using this novel model the present study was aimed to characterize the cell types that specifically respond toward MADB106 tumor cells in lungs of F344 rats within the initial 5-360 min after i.v. inoculation either under intact or under NK cell depleted conditions and to compare these changes with either sham (saline) or splenocyte control injections.…”

mentioning

confidence: 99%

Kinetics of the early recruitment of leukocyte subsets at the sites of tumor cells in the lungs: Natural killer (NK) cells rapidly attract monocytes but not lymphocytes in the surveillance of micrometastasis

Shingu

Helfritz

Kuhlmann

et al. 2002

Intl Journal of Cancer

Self Cite

View full text Add to dashboard Cite

Key words: Fischer 344 rat; MADB106 adenocarcinoma; lung metastasis; natural killer (NK) cell; NK cell depletion; monocytes; Tlymphocytes; immunohistochemistryThe initial sequence and early kinetics within minutes and hours of host defense mechanism against metastatic cells are widely unknown. Leukocyte recruitment at sites of tumor is presently investigated in a time frame of several hours and days. 1 A better understanding of the processes and the cellular components constituting the very early first line of tumor defense in vivo/in situ is needed to unravel the mechanisms of clearance and control of organ metastases.Within hours, tumor cells are rapidly "trapped" to their target organs, such as the lungs. During these early time points a nonspecific "clearance rate" of i.v. injected cells from the lungs has originally been shown by Fidler 2 and since reproduced by others. 3,4 These data indicate that during the first several hours after i.v. injection of tumor cells, the lungs clear themselves from non-specifically trapped tumor cells, until at Day 2-5 preferently those tumor cells remain in the lungs that are suspected to produce lung metastases. 2,5 The rate of clearance, however, seems to be increased when tumor cell-binding endothelial cell adhesion molecules are blocked with adhesion-inhibiting antibodies, indicating a close relation between clearance and the expression of particular endothelial cell adhesion molecules. 1 In addition to these functional effects of cell adhesion molecules in the initial "non-specific" tumor cell trapping, also cell-mediated processes might play a role in determining the initial clearance rate of target organs. At least for natural killer (NK) cell dependent tumor models such as the B16 and the MADB106 cell lines a close relationship between NK cell activity exclusively during the initial 24 hr of host defense and the final disease outcome has been demonstrated. At least 70% of MADB106 tumor cells are destroyed in vivo during the first 1-5 hr after injection 4,6 illustrating that the early host defense response against this tumor cell line is crucial. Recent studies demonstrate that stress hormones such as adrenaline modulate the lung retention of MADB106 tumor cells even within minutes. 7 Thus, these data suggest that detailed time kinetic studies of the cellular changes within the first minutes and hours will provide new insights into specific mechanism determining the initial clearance rate of tumor cells in target organs and for the interaction of different leukocyte subsets in the early surveillance of metastasis.To monitor rapid and specific cellular changes within the lungs as a target organ for metastases detailed studies on the interaction of tumor cells with immune cells in situ were required. The present approach combines immunohistochemic methods with vital dye labeling techniques and takes advantage of the visualization and quantification of effector-like cells that co-localized with CFSE (5-(and -6)-carboxyfluorescein diacetate succinimidyl ester) positive MA...

show abstract

“…The CFSE allows analysis of cell division and precursor frequency both in vivo and in vitro (28,29,(37)(38)(39). One can track T cell proliferation using CFSE fluorescent dye where fluorescence halves in daughter cells of each cell division (40).…”

mentioning

confidence: 99%

Long-Term Cardiac Allograft Survival across an MHC Mismatch after “Pruning” of Alloreactive CD4 T Cells

Watson

Zhang

et al. 2008

The Journal of Immunology

View full text Add to dashboard Cite

Specific tolerance to allografts has been achieved by a variety of means. We have previously shown that ex vivo removal of dividing CD4+ T cells from an MLR or “pruning” delays skin allograft rejection. We tested pruning of alloreactive T cells as a strategy for retaining a broad T cell repertoire while removing alloreactive T cells in a model of cardiac allograft transplant. Using CFSE staining of responder BALB/c cells with stimulator C57BL/6 cells in an MLR, SCID mice were reconstituted with either dividing (D) or nondividing (ND) CD4+ T cells derived from an MLR and then challenged with heterotopic cardiac allografts. Mice reconstituted with D CD4+ T cells rejected cardiac allografts from the stimulator strain with a median survival time (MST) of 29 days, while mice reconstituted with ND CD4+ T cells maintained allografts from the stimulator strain (MST of >100 days) while rejecting third-party allografts (B10.BR) (MST = 11 days). ELISPOT assays demonstrate donor-specific hyporesponsiveness of the ND CD4+ T cells. TCR β-chain V region (TRBV) repertoire analysis demonstrates clonal expansion within both rejecting D cardiac allografts and ND cardiac allografts surviving for the long-term. Histology showed greater allograft infiltration by the D CD4+ T cells. The surviving ND cardiac allografts demonstrated reduced cellular infiltration and reduced incidence of allograft vasculopathy, but with the development of chronic fibrosis. Thus, pruning of alloreactive T cells allows long-term-specific cardiac allograft survival while retaining the ability to reject third-party allografts.

show abstract

Stereological quantification of carboxyfluorescein-labeled rat lung metastasis: a new method for the assessment of natural killer cell activity and tumor adhesion in vivo and in situ

Cited by 26 publications

References 33 publications

Diurnal changes in lung tumor clearance and their relation to NK cell cytotoxicity in the blood and spleen

Diurnal changes in lung tumor clearance and their relation to NK cell cytotoxicity in the blood and spleen

Kinetics of the early recruitment of leukocyte subsets at the sites of tumor cells in the lungs: Natural killer (NK) cells rapidly attract monocytes but not lymphocytes in the surveillance of micrometastasis

Long-Term Cardiac Allograft Survival across an MHC Mismatch after “Pruning” of Alloreactive CD4 T Cells

Contact Info

Product

Resources

About