• IHC/IF/ICC targets classification:
  • Lymphocyte lineage: CD3, CD4, CD8, CD19, CD20, CD27, CD38, CD45, CD47, CD69, CD79a, CD103, CD161, PD-1, FOXP3, OX40, LAG3, 4-1BB, NK1.1, NCR1, TCR, MHC-1, FCRn, etc.
  • Myeloid lineage : CD68, CD86, CD163, CD206, iNOS, F4/80, CD45, CD38, PD-1, CD11c, CD11b, Ly6c, Ly6g, tryptase, etc.
  • TAA / TSA: B7H3, p53, CK, CK5, TOP2, TROP2, HER2, HER3, ER, PR, PD-L1 /PD-1, EGFR, Claudin18.2, MUC1, TPBG, ROR1, LILRB4, E Cadherin, WT1, PTK7, EPCAM, MET, GPC1, MUC16, DLL3, etc.
  • Cytokines / cytokine receptors: IFN-γ, IL23, Glucagon, Insulin, CXCR1, TGF beta Receptor, CCR3, CCR8/CD198, etc.
  • Micrangium / blood-brain barrier: CD31, VEGF Receptor2, TFR1, IGF1R, CD98h, LAT1, etc.
  • Phosphorylation : c-MET, p-ERK, etc.
  • Others: C3/C3a, ASPH, ZP2, β-Amyloid (1-40), β-Amyloid (1-42), MED13L, uPAR, etc.
• RNA scope: UBC, IL23, IL4, IL13, etc.
• FISH/ISH: c-MET, EGFR, etc.

Pembrolizumab demonstrated a significant anti-tumour effect in a dose-dependent manner in MC38 xenograft model in B-hPD-1 mice. Reduced PD-1 expression in macrophages was seen in G4 compared to control, indicating macrophages are critical effector cells in anti-PD-1 treatment.

An increase of M1 was found in invasive margin of MC38 tumors, whereas M2 declined significantly in response to pembrolizumab treatment, with a dose-dependent manner. This indicates M1 infiltration facilitated anti-PD-1 mediated tumor growth inhibition.

TMA staining shows that CD8+ T (CTL) cells were increased dramatically in both tumor invasive margins and central tumor areas of Test. This is associated with an increased M1/M2 ratio.

Main features of Asthma:

• Chronic inflammation of the respiratory tract: mixed inflammatory cell infiltration around the bronchus and blood vessels
• Respiratory tract hyperresponsiveness and airflow obstruction: mucus formation in the bronchus

Pathological analysis available:

• Diagnosis of lesions, to judge whether the modeling is successful
• Count the differences between groups and analyze the pros and cons of different model-building doses, methods, animals, etc.
• Based on the pathological results, make reasonable suggestions for the next test
• Combined with drug pharmacology, analyze the relieving effect of different drugs on the disease and the difference in drug effect

HE staining- inflammation in psoriasis skin tissue

HE staining- inflammation in atopic dermatitis skin tissue

Main features of Non-alcoholic steatohepatitis (NASH):
NASH is a form of NAFLD in which has hepatitis inflammation of the liver and liver cell damage, in addition to fat in the liver. Inflammation and liver cell damage can cause fibrosis, or scarring of the liver. NASH may also lead to cirrhosis or liver cancer.

Immunohistochemistry showing fibroblast marker α-SMA in liver

Main features of IBD:

• Tissue destruction: loss of intestinal gland, depletion of goblet cells, and formation of ulcer in mucosal layer
• Intestinal inflammation: edema of mucosal layer/ submucosa/ muscularis with mixed inflammatory cell infiltration

Note: No significant pathological changes were observed in the colon of control group animals microscopically. In the DSS-induced model group, the colon showed crypt loss, ulcer formation, and infiltration of mixed inflammatory cells. Black arrows indicate crypt loss, black triangles indicate ulcer formation in the mucosal layer, and black stars indicate inflammatory cell infiltration.

Note: No significant pathological changes were observed in the colon of control group animals microscopically. In the DSS-induced model group, fibrous proliferation was observed in the colon. Black arrows indicate fibrous proliferation.

Note: No significant pathological changes were observed in the colon of control group animals microscopically following 40% ethanol instillation. In the TNBS-induced model group, ulcer formation, transmural inflammation, and intestinal necrosis were observed in the colon. Black arrows indicate ulcer formation in the mucosal layer, black triangles indicate intestinal necrosis, and black stars indicate transmural inflammation.

Note: No significant pathological changes were observed in the colon of FACS control group animals microscopically. In the CD4+CD25– adoptive transfer model group, crypt loss and inflammatory cell infiltration were observed in the colon. Black arrows indicate crypt loss, and black stars indicate inflammatory cell infiltration.

Main features of CIA:
CIA is the most widely studied and used standard model, among the various animal models of rheumatoid arthritis (RA). The pathological features CIA model shared with RA including a proliferative synovitis with infiltration of polymorphonuclear and mononuclear cells, cartilage destruction, bone resorption, pannus formation and fibrosis.

HE staining: inflammatory cell infiltration, cartilage destruction, bone resorption and pannus formation

Main features of Surgically-Induced OA Models:
Surgical methods of OA induction work through a combination of joint destabilization, altered joint mechanics, and intra-articular inflammation. Anterior Cruciate Ligament Transection (ACLT )model is induced due to ACL injury, which causes joint destabilization, subsequently leading to Post-traumatic osteoarthritis (PTOA). The ACLT-induced OA model in mouse has many features such as cartilage destruction, osteophyte formation and synovial changes.

Saffron-O and Fast Green staining-cartilage destruction

Preclinical toxicology studies are largely focused on drug safety evaluation. Whether a drug is safe and effective is the determining factor in drug development, and toxicity/safety is a major cause for study discontinuation throughout the whole drug development process. The aim is to provide a reliable toxicokinetic basis for human clinical trials by studying the relationship between drug exposure and toxic response in animal models, explaining possible target organs of toxicity and toxic responses, and predicting drug safety in humans.

Team Introduction

• Technical Lead:
  • Certified Veterinary Pathologist (China) and Member of the Japanese Society of Toxicologic Pathology
• Extensive Experience:
  • Over 10 years in toxicologic pathology and more than 6 years in efficacy pathology
  • Successfully managed over 10 dual-filing projects
• Pathology Expertise:
  • Toxicologic pathology experience with vaccines, antibodies, small molecules, and oligonucleotide drugs
  • Animal models: Cynomolgus monkeys, beagle dogs, rats, mice, rabbits, and miniature pigs
  • Processed and analyzed over 100,000 tissue sections
• Project Pathology Lead:
  • Over 4 years of project leadership in pathology
  • Reviewed more than 30,000 slides across various projects

Increased acute phase proteins and liver damage in response to Selicrelumab (analog) in humanized B-hCD40 mice

Anaemia and reticulocytosis in humanized B-hSIRPa/hCD47 mice

RBC and Hemoglobin are transiently reduced at the early stage of high dose Hu5F9 treatment and return to normal levels.

Hyperbilirubinemia, lymphocyte count decreased and chills in humanized B-hSIRPa/hCD47 mice