drcsukasdomokos@gmail.com This research was supported by

the TheraGlio EU Project: 602923.

Pulmonary intravascular macrophages: Prime suspects as cellular triggers of porcine CARPA

Domokos Csukás1, Rudolf Urbanics2, György Wéber1, László Rosivall3-4, János Szebeni2-5

1Department of Surgical Research and Techniques, Semmelweis University, Budapest, Hungary 2Seroscience Ltd., Budapest, Hungary

3Department of Pathophysiology, International Nephrology Research and Training Center, Semmelweis University, Budapest, Hungary 4Nanomedicine Research and Education Center, Semmelweis University, Budapest, Hungary

5Department of Nanobiotechnology and Regenerative Medicine, Faculty of Health, Miskolc University, Miskolc, Hungary

Csukás D. et al. : Pulmonary intravascular macrophages: prime suspects as cellular mediators of porcine CARPA , Eur. J. Nanomed. 2015; 7(1): 27–36, DOI 10.1515/ejnm-2015-0008

Credits:CDC , E. Ewing, J. Hedberg, wiki, Ruoff Gr.,Worldchoice, Urbanics, Szebeni

100 nm

DOXIL®/Caelyx® HIV-1

1nm=10-9 m 10nm 100 nm 1000 nm = 1 μm

C60-Fullerene Micelle Liposome Carbon nanotube

NANODRUGS = THE TRIGGER

• VARIOUS STRUCTURES AND FUNCTION

• SIMILAR TO VIRUSES IN SIZE & SHAPE

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SAP mean

HR mean

PAP, SAP and HR % changes after repeated liposomal injections

% v

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•COMPLEX CARDIO-PULMONARY

SYMPTOMS, SIMILAR TO HUMAN PATIENTS

• UNIQUELY HIGH SENSITIVITY

PORCINE MODEL & REACTION

Human appearance and effect of PIMs? • In hepatopulmonary syndrome macrophage accumulation in the

intravascular space of the lung is present • Certain healthy people may have PIMs

Conclusions: Ex vivo examinations, the details of the role of PIMs, and the possibility of selective depletion may present a potential therapeutic target preventing CARPA.

IS THERE A CELLULAR CONTRIBUTOR OF CARPA IN THE CARDIOPULMONARY TRACT?

PULMONARY INTRAVASCULAR MACROPHAGES (PIMs)

Morphology Porcine PIMs • PIM is reaching out with pseudopods

attached to the surface of plastic coverslip after incubation (sizebar 2 μm);

• PIMs showing rounded shape, ruffled membrane after incubation on the pulmonary artery endothelium ex vivo, (sizebar 5 μm).

Modified from Morton et al. J Leukocyte Biol 1988 with permission

Methodology Porcine PIM cells adhered to plastic surface. The dens, larger cells (marked by arrows) show ruffled membrane structures and multiple vacuoles. Cells were washed out from the capillaries by collagenase and let to adhere to gelatin surface , 40 × .

Strong adherence of PIM cells enables their in vitro separation. Csukás, et al. Eur. J. Nanomed. 2015

Adherence and Phagocytosis PIM of a sheep, anchored to the pulmonary capillary endothelium via junction-like intercellular adhesion plaques (ICAPs). Characteristic features include indented nucleus and a unique glycocalyx membrane. This surface plays a key role in receptor–mediated clearance of phagocytosis . Unique ruffled globular membrane structure (arrows). AS, alveolar space; *, red blood cells; circle, phagocytosis; thick arrow, capillary wall; curved arrow, alveolar epithelium; magnification, X10,000. From Schneberger et al. Am J Physiol Lung Cell Mol Physiol, 2012.

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TxB2 & PAP changes after Zymosan bolus injection (0.5 mg/kg)

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PAP mean (mmHg)

Secretion and Mediators Time correlation between the rise of blood TxB2 and pulmonary arterial pressure during (zymosan-induced) CARPA in a pig. They also express patternrecognising and anaphylatoxin receptors on their surface, this way C activation can trigger these cells. Szebeni J. Mol Immunol 2014.

CARPA Complement Activation-related Pseudoallergy

TYPE I hypersenzitivity reaction without IgE

Caused by wide range of agents

Reactions from mild and reversable to lethal

CLINICAL and REGULATORY SIGNIFICANCE

Presented at 2015 Conference

Appearance • Animal species with RESIDENT PIMs from

birth: pig, sheep, cattle, horse, cat,etc. • Species with PIMs appear after

INDUCTION of foreign agents: Rodents, Rabbit, Dogs, Macaques…Humans?

Brain et al. Am J Physiol Lung Cell Mol Physiol 1999

.