Rapid quantification of lymphocyte subsets in heterogeneous cell populations by flow cytometry

Pechhold, Klaus; Pohl, Thomas; Kabelitz, Dieter

doi:10.1002/cyto.990160209

Cited by 95 publications

(67 citation statements)

References 8 publications

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“…Briefly, total cell numbers in individual HKL isolates were determined by means of flow cytometry with the standard cell dilution assay (Pechhold et al 1994) as modified by Scharsack et al (2004). Suspensions of HKL were adjusted to 1.2!10 5 viable cells per millilitre with RPMI 1640 diluted with 10% (v/v) distilled water (R-90).…”

Section: Methodsmentioning

confidence: 99%

Who is in control of the stickleback immune system: interactions between Schistocephalus solidus and its specific vertebrate host

2007

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The cestode Schistocephalus solidus is a frequent parasite of three-spined sticklebacks and has a large impact on its host's fitness. Selection pressure should therefore be high on stickleback defence mechanisms, like an efficient immune system, and also on parasite strategies to overcome these. Even though there are indications for manipulation of the immune system of its specific second intermediate host by the cestode, nothing is yet known about the chronology of specific interactions of S. solidus with the stickleback immune system. We here expected sticklebacks to first mount an innate immune response directly post-exposure to the parasite to clear the infection at an early stage and after an initial lag phase to upregulate adaptive immunity. Most interestingly, we did not find any upregulation of the specific lymphocyte-mediated immune response. Also, the pattern of activation of the innate immune system did not match our expectations: the proliferation of monocytes followed fluctuating kinetics suggesting that the parasite repeatedly installs a new surface coat not immunogenic to the host. Furthermore, the respiratory burst activity, which has the potential to clear an early S. solidus infection, was upregulated very late during infection, when the parasite was too big to be cleared but ready for transmission to its final host. We here suggest that the late activation of the innate immune system interferes with the neuroendocrine system, which mediates reduced predation avoidance behaviour and so facilitates the transmission to the final host.

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Section: Methodsmentioning

confidence: 99%

Who is in control of the stickleback immune system: interactions between Schistocephalus solidus and its specific vertebrate host

2007

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“…Total numbers of cells were recorded by means of the standard cell-dilution assay (Pechhold et al 1994). Culture plates were placed on ice for 10 min, shaken, and the whole content of each well was transferred to individual flow cytometer tubes.…”

Section: Fishmentioning

confidence: 99%

Effect of hexavalent carcinogenic chromium on carp Cyprinus carpio immune cells

Steinhagen¹,

Helmus²,

Maurer³

et al. 2004

Dis. Aquat. Org.

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“…The quantification of cultured cells was performed according to the standard cell dilution method (Pechhold et al 1994). Known numbers of standard cells (2 X 105) were added to each tube with cultured cells.…”

Section: Methodsmentioning

confidence: 99%

Flow cytometric analysis of proliferative responses of carp Cyprinus carpio peripheral blood leukocytes to mitogens and to the hemoflagellate Trypanoplasma borreli

Scharsack¹,

Steinhagen²,

Körting³

et al. 2000

Dis. Aquat. Org.

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The activation of carp peripheral blood leukocytes (PBL) was analysed radiometrically and by means of flow cytometry (FCM) in order to compare the results obtained with both methods. The qualitative and quantitative FCM analyses of cellular morphology and viability resulted in a further characterisation of proliferative responses of carp PBL to Trypanoplasma borreli in vivo and in vitro. The lymphocyte population of PBL from T borreli-infected carp exhibited a marked shift in forward scattered light (FSC; cell size). When PBL from healthy carp were stimulated with mitogens in vitro, a lymphoid population with increased FSC profiles was also observed. The number of these cells coincided to ratios of 3H-thymidine incorporation, recorded from corresponding cultures. Thus, it was concluded that the increase in size of stimulated lymphocytes could be due to blastogenic transformation. The advantage of the FCM procedure is that activation and proliferation of carp lymphocytes can be monitored without labelling the cells. Cocultures of mitogen-stimulated carp PBL and T borreli revealed the ability of the parasite to suppress lymphocyte proliferation in vitro.

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Rapid quantification of lymphocyte subsets in heterogeneous cell populations by flow cytometry

Cited by 95 publications

References 8 publications

Who is in control of the stickleback immune system: interactions between Schistocephalus solidus and its specific vertebrate host

Who is in control of the stickleback immune system: interactions between Schistocephalus solidus and its specific vertebrate host

Effect of hexavalent carcinogenic chromium on carp Cyprinus carpio immune cells

Flow cytometric analysis of proliferative responses of carp Cyprinus carpio peripheral blood leukocytes to mitogens and to the hemoflagellate Trypanoplasma borreli

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