Table 3.

Overview of level 3 preclinical studies according to the search criteria

First Author	Year	Tumor type	Implantation site	Animal	Immune competent /Syngeneic	Radiotherapy (Site – Total Dose [Gy] / Fractions)	Immunotherapy	Observed Effect	Level of response	Suggested mediator
Buchegger25	1995	Col 12 and LS174T	Transplants on the middle of their backs at 2 cm from the tail	nude mice	- / +	Back – 16 Gy/2x	3 monoclonal anti-CEA antibodies (mAb 35, CE25-B7, and B93)	Tumor growth delay	3	NA
Chiang26	2000	Fibrosarcoma – C3H/HeN	s.c. in the right thigh	C3H/HeJ mice	+ / +	right thigh – 25 to 35 Gy/1 x	IL-3 tumor vaccin	Tumor growth delay	3	increased intra tumoral levels of intercellular adhesion molecule-1, Mac-1, EB22/5.3, tumor necrosis factor
Lohr27	2000	4T1 mammary tumor cfC3H – B16 melanoma	s.c. in the right hind leg	BALB + C57BL/6 mice	+ / +	Leg – 18 to 33 Gy/3 x	Adenovirus, IL-12 – B7.1	Tumor growth delay	3	Upregulation T-cells and NK-cells
Teitz- Tennenbaum28	2003	D5melanoma or MCA 205 sarcoma	s.c. in the mid-right flank	C57BL/6 mice	+ / +	Flank – 42.5 Gy/5x	Dendritic cell	D5: Tumor inhibition 65.9% MCA 205: tumor inhibition	3	IFN-ℽ production by host-derived T cells
Huang29	2007	renal cell carcinoma	s.c. in the right axilla	C57Bl/6 mice	+ / +	Right Axilla – 35 Gy / 5x	Dendritic cell	Tumor growth delay	3	Down-regulation of Bcl-2, up-regulation of Bax, Expression of TNFα, IL-2, IL-4 m IFN-γ, IgG, IgM.
Meng30	2012	melanoma cell line B16SIY	s.c. in the right leg	C57Bl/6 mice	+ / +	Right Leg – 6 or 12 Gy / 1x	poly(ADP-ribose) polymerase inhibitor veliparib	Tumor growth delay	3	express immune stimulatory cytokines (CCL2, CCL5, CXCL9, CXCL10, and CXCL11) to activate cytotoxic T lymphocytes
Wang31	2012	T-26, a murine colon carcinoma cell line	s.c.in the left flank	BALB/c mice	+/+	Left flank – 8 Gy / 1x	Dendritic cell + Recombinant heat shock protein 70	Tumor growth delay	3	Expression of TNFα, IL-12, T-cell upregulation
Wei32	2013	murine D5 melanoma	s.c.in the right flank	C57BL/6 (B6) and B6.PL-Thy1a/CyJ (CD90.1) mice	+/+	bilateral flanks – 8.5 Gy / 5x	IL-2	Tumor growth delay	3	expression of the effector cytokines IFN-γ and TNF-α by donor and host CD4+ and CD8 + T cells
Dovedi33	2014	CT26 murine colon carcinoma cells	s.c. – not further specified	BALB/c and C57Bl/6 mice	+ / +	Local – 10 Gy / 5x	anti PD-L1	Tumor growth delay	3	NK cells, CD8+ T-cell upregulation
Lim34	2014	B16 melanoma	i.m. into the lower left thigh	C57BL/6J	+ / +	Leg – 15 Gy / 1x	Listeria monocytogenes-based cancer vaccine	Tumor growth delay	3	increase in intratumoral numbers of activated T cells, antigen-specific CD8 + t cells, natural killer cells + levels of effector molecules, such as interferon γ and granzyme B
Rekers35	2015	F9 terato-carcinoma cells	s.c.in the flank	129/SvHsd mice		Flank – 12 Gy /1x	L19-IL2	Tumor growth delay	3	Extra Domain-B expression and infiltration of cytotoxic T cells.
Blanchard36	2015	B16-OVA melanoma	s.c. in the hind limb.	C57BL/6 mice	+ / +	Lower limb – 20 Gy/1x	vesicular stomatitis virus- tumor-associated antigen viral immunotherapy	Tumor growth delay	3	Upregulation T cells
Mondini37	2015	TC1/Luccells / HNSCC implantation model –	submucosal site of the right inner lip	C57BL/6 mice	+ / +	head and neck region – 2.6 −7.5 Gy /1-4x	STxB- E7vaccine	Tumor growth delay	3	tumor-infiltrating, antigen-specific CD8 + T cells
Sharabi38	2015	MC38-OVA cells; B16-OVA melanoma cells; 4T1HA breast carcinoma cell	s.c.in the right flank	C57BL/6, BALB/cJ, and MHC Class I knockout mices	+/+	Right flank – 10 to 20 Gy / 1x	Anti- PD-L1	Tumor growth delay	3	increased T-cell infiltration in tumor: CD8+, CD4+ CD25+ Foxp3 + T-regulatory cells
Monjazeb39	2016	B16 melanoma or 4T1 breast adenocarci-noma	into the flank	C57BL/6 or BALB/c mice	+ / +	Mammary fat pad / flank – 8 Gy/ 1x	CpG oligodeoxynucleotide, enzyme indolamine-2,3-dioxygenase blockade	Tumor growth delay	3	upregulation Toll-like Receptor 9, CD4+
Young40	2016	CT26 murine colorectal carcinoma	s.c.in the right hind limb	BALB/c and FVB mice	+/+	Limb – 20 Gy / 1x	OX40 (CD134) Anti-CTLA4	Tumor growth delay	3	Depletion of CD4+ or CD25
Zheng41	2016	Panc02 and MC57-SIY cells	s.c. on the back of the mice	C57BL/6 mice	+ / +	Local – 20 Gy/1x	Vaccination+ antiPD-L1	Tumor growth delay	3	CD8 + T cell infiltration; upregulation of CXCL10 and CCL5 chemokine
Oweida42	2017	LY2 + B4B8 squamous cell carcinoma	Submucosal via the buccal mucosa	BALB/c mice	+/+	Buccal + regional neck level – 10 Gy/1x	anti PD-L1	Tumor growth delay	3	upregulation of PD-L1 increased T-cell infiltration in tumor
Weiss43	2017	SMA glioma cell lines	intracranial (the right striatum)	C57BL/6 CD45.2 mice	+/+	Cranial – 4 Gy/1x	NKG2D-Based CAR T Cells	Tumor growth delay	3	high IFNg production and cytolytic activity in vitro
Choi44	2018	CT-26 colon carcinoma cells	s.c. into the right legs and left flanks	BALB/c mice	+/+	Leg – 15 Gy/1x –	Dendritic cell	Tumor growth delay	3	Maximum Dendritic cell sensitization and T-cell stimulation with IL-10, IL-12, and interferon (IFN)-γ production
Wang45–50	2019	Lewis lung carcinoma	s.c. into the left upper flank	C57Bl/6 mice	+ / +	Flank – 24 Gy/3x	α-PD-L1	Tumor growth delay	3	CD8 T-cell infiltration; PD-L1 expression

s.c.: subcutaneous, i.m.: inta-musculair, PR: Partial Response, NA: not appropriated