Current Research topics
- Development of scientific and clinical insights into the concept of 'Split
Immunity'. The concept is soon to be translated from bench to bedside.
- Mapping of the entire adaptive and innate cellular immune milieu found
inside human brain tumors using advanced multicolor (up to 8-color) flow
- Using an approach termed "immune cytomics" that uses systems-biology and a
wet-immunology, to study the network of intra-tumoral cellular interactions.
Such interactions are formed between the different subsets of immune cells, and
between immune cells and tumor cells.
- Development of a high-throughput proteomic- genomic-immunologic approach to
identify the immunodominant tumor associated antigens recognized by tumor
infiltrating T cells.
- Identification of systems-level predictive and prognostic markers for
detection of effective anti-tumor responses in glioma patients.
Animal models for brain tumors:
We use stereotactic injections to inoculate rats and mice with several
different types of syngeneic ('autologous') brain tumors. These brain tumor
models are highly aggressive and closely resemble brain tumors found in humans.
Additionally we have developed a rat glioma cell-line that expresses luciferase.
This cell-line enables the in-vivo monitoring of the quantity of living cells
that grow inside a rat's brain, using the IVIS system.
Tumor enzymatic dissociation to single cells:
We developed a method to enzymatically dissociate human and animal brain
tumors to single cells with very high viability (>90%). Working with ex-vivo
produced cells of high viability reduces the frequent in-vitro artifacts caused
by the excessive death of cells dissociated by other methods.
Advanced flow cytometry:
We developed several different flow-cytometric antibody panels to identify
all prominent immune cells found inside a brain tumor . Using these carefully
designed multicolor (~8 color) panels we can currently identify all tumor
infiltrating and tumor resident immune cells found within human brain tumors,
including: CTL, Helper T cells, NK cells, NKT cells (2 subsets), macrophages
(M1/M2), plasmacytoid dendritic cells, myeloid dendritic cells (3
subtypes),neutrophils, eosinophils, and microglia (2 activation states). The
correct identification of these cells enables their purification via sorting.
Purified cells are then used in order to run subsequent high-resolution genomic
or proteomic assays.
Functional ex-vivo assays:
Short (1-3d) ex-vivo assays are done on freshly isolated tumor and
peripheral blood leukocytes. Such assays may uncover subtle immune responses on
the single cell-level of either peripheral immune cells or intratumoral immune
We use these functional flow cytometric panels to follow immune
functional markers such as: TNF-alpha, TGF-beta, IFN-gamma, Interleukins (e.g.
IL2, IL13), CD107a (cytotoxicity/degranulation marker), PD1 (T-cell
activation/exhaustion marker), proliferation (CFSE), and cellular viability (via
amine dyes). Each of the constructed T-cell functional panels also follows
T-cell identity (using CD3/4/8/(DUMP: CD14- 19- 20- Viability) ) and four of
the functional markers listed above.
In addition to T-cell functional
panels we have also constructed several panels to functionally monitor human
dendritic cells. These panels follow the activation and maturation state of all
four subtypes of human dendritic cells in peripheral blood and within the
• ICRF - Israel Cancer Research Fund grant (current
• VABC - Voices against brain cancer grant
• ICA - Israel Cancer Association grant
• MOH - Israeli Ministry of Health grant
- The Dept of Immunology – the Weizmann Institute of Science.
- Advanced technologies center – Sheba Medical Center
- Koret School of Veterinary Medicine - The Hebrew University, Beit
Nati Shapira, MSc - researcher
Merav Bloch, MSc
Tal Alter, MSc
Idan Ben Horin, MD
Ori Barzilay, MD
Anatoly Shinkarevsky, MSc student