Biochemical Analysis of Leukocytes after In Vitro and In Vivo Activation with Bacterial and Fungal Pathogens Using Raman Spectroscopy

Pistiki, Aikaterini; Ramoji, Anuradha; Ryabchykov, Oleg; Thomas-Rüddel, Daniel; Press, Adrian T.; Makarewicz, Oliwia; Giamarellos-Bourboulis, Evangelos J.; Bauer, Michael; Bocklitz, Thomas; Popp, Jürgen; Neugebauer, Ute

doi:10.3390/ijms221910481

Open AccessArticle

Biochemical Analysis of Leukocytes after In Vitro and In Vivo Activation with Bacterial and Fungal Pathogens Using Raman Spectroscopy

by

Aikaterini Pistiki

^1,2,†,

Anuradha Ramoji

^1,2,3,†

,

Oleg Ryabchykov

^1,3

,

Daniel Thomas-Rüddel

^2,4

,

Adrian T. Press

^2,4,5

,

Oliwia Makarewicz

⁶,

Evangelos J. Giamarellos-Bourboulis

⁷,

Michael Bauer

^2,4,8,

Thomas Bocklitz

^1,3

,

Jürgen Popp

^1,2,3,8

and

Ute Neugebauer

^1,2,3,8,*

¹

Leibniz Institute of Photonic Technology Jena (a Member of Leibniz Health Technologies), Albert-Einstein-Straße 9, 07745 Jena, Germany

²

Center for Sepsis Control and Care, Jena University Hospital, Am Klinikum 1, 07747 Jena, Germany

³

Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich Schiller University, Helmholtzweg 4, 07743 Jena, Germany

⁴

Department of Anesthesiology and Intensive Care, Jena University Hospital, Am Klinikum 1, 07747 Jena, Germany

⁵

Medical Faculty, Friedrich Schiller University, Bachstraße. 18, 07743 Jena, Germany

⁶

Institute of Infectious Diseases and Infection Control, Jena University Hospital, Am Klinikum 1, 07747 Jena, Germany

⁷

4th Department of Internal Medicine, Medical School, National and Kapodistrian University of Athens, Rimini Street 1, 12462 Athens, Greece

⁸

Jena Biophotonics and Imaging Laboratory, Albert-Einstein-Straße 9, 07745 Jena, Germany

^*

Author to whom correspondence should be addressed.

^†

These authors contributed equally to this work.

Int. J. Mol. Sci. 2021, 22(19), 10481; https://doi.org/10.3390/ijms221910481

Submission received: 29 July 2021 / Revised: 14 September 2021 / Accepted: 23 September 2021 / Published: 28 September 2021

(This article belongs to the Special Issue Infrared and Raman Spectroscopy of Human Diseases)

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Abstract

Biochemical information from activated leukocytes provide valuable diagnostic information. In this study, Raman spectroscopy was applied as a label-free analytical technique to characterize the activation pattern of leukocyte subpopulations in an in vitro infection model. Neutrophils, monocytes, and lymphocytes were isolated from healthy volunteers and stimulated with heat-inactivated clinical isolates of Candida albicans, Staphylococcus aureus, and Klebsiella pneumoniae. Binary classification models could identify the presence of infection for monocytes and lymphocytes, classify the type of infection as bacterial or fungal for neutrophils, monocytes, and lymphocytes and distinguish the cause of infection as Gram-negative or Gram-positive bacteria in the monocyte subpopulation. Changes in single-cell Raman spectra, upon leukocyte stimulation, can be explained with biochemical changes due to the leukocyte’s specific reaction to each type of pathogen. Raman spectra of leukocytes from the in vitro infection model were compared with spectra from leukocytes of patients with infection (DRKS-ID: DRKS00006265) with the same pathogen groups, and a good agreement was revealed. Our study elucidates the potential of Raman spectroscopy-based single-cell analysis for the differentiation of circulating leukocyte subtypes and identification of the infection by probing the molecular phenotype of those cells.

Keywords: Raman microspectroscopy; leukocytes; infection model; Staphylococcus aureus; Klebsiella pneumoniae; Candida albicans; neutrophil; PBMC; monocyte; lymphocyte

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MDPI and ACS Style

Pistiki, A.; Ramoji, A.; Ryabchykov, O.; Thomas-Rüddel, D.; Press, A.T.; Makarewicz, O.; Giamarellos-Bourboulis, E.J.; Bauer, M.; Bocklitz, T.; Popp, J.; et al. Biochemical Analysis of Leukocytes after In Vitro and In Vivo Activation with Bacterial and Fungal Pathogens Using Raman Spectroscopy. Int. J. Mol. Sci. 2021, 22, 10481. https://doi.org/10.3390/ijms221910481

AMA Style

Pistiki A, Ramoji A, Ryabchykov O, Thomas-Rüddel D, Press AT, Makarewicz O, Giamarellos-Bourboulis EJ, Bauer M, Bocklitz T, Popp J, et al. Biochemical Analysis of Leukocytes after In Vitro and In Vivo Activation with Bacterial and Fungal Pathogens Using Raman Spectroscopy. International Journal of Molecular Sciences. 2021; 22(19):10481. https://doi.org/10.3390/ijms221910481

Chicago/Turabian Style

Pistiki, Aikaterini, Anuradha Ramoji, Oleg Ryabchykov, Daniel Thomas-Rüddel, Adrian T. Press, Oliwia Makarewicz, Evangelos J. Giamarellos-Bourboulis, Michael Bauer, Thomas Bocklitz, Jürgen Popp, and et al. 2021. "Biochemical Analysis of Leukocytes after In Vitro and In Vivo Activation with Bacterial and Fungal Pathogens Using Raman Spectroscopy" International Journal of Molecular Sciences 22, no. 19: 10481. https://doi.org/10.3390/ijms221910481

APA Style

Pistiki, A., Ramoji, A., Ryabchykov, O., Thomas-Rüddel, D., Press, A. T., Makarewicz, O., Giamarellos-Bourboulis, E. J., Bauer, M., Bocklitz, T., Popp, J., & Neugebauer, U. (2021). Biochemical Analysis of Leukocytes after In Vitro and In Vivo Activation with Bacterial and Fungal Pathogens Using Raman Spectroscopy. International Journal of Molecular Sciences, 22(19), 10481. https://doi.org/10.3390/ijms221910481

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Biochemical Analysis of Leukocytes after In Vitro and In Vivo Activation with Bacterial and Fungal Pathogens Using Raman Spectroscopy

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