Co-culture or tumour hepatocytes with homologous peripheral lymphocytes in vitro and study of their interaction

Doctoral Dissertation uoadl:1305611 304 Read counter

Τομέας Χειρουργικής
Library of the School of Health Sciences
Deposit date:
Δούμπα Πολυξένη
Dissertation committee:
Ιωάννης Μπράμης, Ιωάννης Κοσκίνας (επιβλέπων), Μανούσος Κωνσταντουλάκης
Original Title:
Αλληλεπίδραση της συγκαλλιέργειας καρκινικών ηπατοκυττάρων με ομόλογα περιφερικά λεμφοκύτταρα in vitro
Translated title:
Co-culture or tumour hepatocytes with homologous peripheral lymphocytes in vitro and study of their interaction
Background: Many studies have suggested that the immune response may play a
crucial role in the progression of hepatocellular carcinoma (HCC). Therefore,
our aim was to establish a (i) functional culture of primary human tumor
hepatocytes and non-tumor from patients with hepatocellular carcinoma (HCC) and
(ii) a co-culture system of HCC and non-HCC hepatocytes with autologous
peripheral blood mononuclear cells (PBMCs) in order to study in vitro
cell-to-cell interactions.
Methods: Tumor (HCC) and non-tumor (non-HCC) hepatocytes were isolated from the
liver resection specimens of 11 patients operated for HCC, while PBMCs were
retrieved immediately prior to surgery. Four biopsies were obtained from
patients with no liver disease who had surgery for non malignant tumor (normal
hepatocytes). Hepatocytes were either cultured alone (monoculture) or
co-cultured with PBMCs. Flow cytometry measurements for MHC class II
expression, apoptosis, necrosis and viability (7AAD) were performed 24 h, 48 h
and 72 h in co-culture and monocultures.
Results: HCC and non-HCC hepatocytes exhibited increased MHC-II expression at
48h and 72h in co-culture with PBMCs as compared to monoculture, with MHC
II-expressing HCC hepatocytes showing increased viability at 72 h. PBMCs showed
increased MHC-II expression (activation) in co-culture with HCC as compared to
non-HCC hepatocytes at all time points. Moreover, CD8+ T cells had
significantly increased apoptosis and necrosis at 48h in co-culture with HCC
hepatocytes as compared to monocultures. Interestingly, MHC-II expression on
both HCC and
non-HCC hepatocytes in co-culture was positively correlated with the respective
activated CD8+ T cells.
Conclusions: We have established an in vitro co-culture model to study
interactions between autologous PBMCs and primary HCC and non-HCC hepatocytes.
This direct interaction leads to increased antigen presenting ability of HCC
hepatocytes, activation of PBMCs with a concomitant apoptosis of activated CD8+
T cells. Although, a partially effective immune response against HCC exists,
still tumor hepatocytes manage to escape.
HCC Hepatocyte-PBMCs Co-Cultures, Non-HCC Hepatocyte-PBMCs Co-Cultures, CD8+ T lymphocyte activation, Apoptosis, MHC-II Expression
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