Table 3 Characteristics of the animal experiments (n = 48)

Animals

Mice

42

Rats (1 × mixed: rats & mice)

4

Horses, Cats

2

Diagnoses

Healthy animals

21

Cancer * (2 × mixed)

27

Treatment

Whole extract

34

Isolated or recombinant ML

14

Application route

Subcutan, intraperitoneal **

35

Intratumoural, intravenous, intramuscular, intracutan, oral, intranasal, intravesical

13

Application frequency

Applied just once

10

Applied more than once (5 × mixed) (up to several months)

38

Maximum dose per application

≤ 20 mg mg/kg VAE

12

> 20-100 mg/kg VAE

16

> 100 mg/kg VAE (maximum dose: 1400 mg/kg)

6

> 50 ng/kg ML (maximum dose: 500 μg/kg oral, 50 μg/kg intranasal, 14 μg/kg sc)

9

< 50 ng/kg ML

5

Observation time < 1 week

6

Treatment of control group

Placebo

33

No additional treatment or unclear

14

No control group

1

Immune outcomes investigated

38

Peripheral blood: CBC, DBC, leukocytes, lymphocytes, monocytes, granulocytes, T-cell subsets, activation markers, TNF-α

14

Immunoglobulins/humoral response to foreign antigens

9

Cellular response to foreign antigens, foreign skin graft rejection

2

Thymus (size, histological analysis, thymocytes, subsets, function

14

Spleen (size, weight, morphometric analysis, splenocytes, subsets, function)

8

Lymph nodes (weight, morphometric analysis)

2

Peritoneal macrophages and activity

3

Influence on leukopenia caused by radiation, chemotherapy, dexamethasone.

4

Others (tumour tissue, urinary bladder tissue, bronchoalveolar lavage)

4

Safety outcomes investigated

17

Monitored for toxicity, tolerability, vitality, clinical signs, body weight, food/water consumption, behavior, physical responses of the animals, local effects

9

Necropsy

2

Bladder histology after intravesical instillation of VAE

1

No details on recording

8

Time schedule of safety assessment

Daily, 2/week

5

„Regular"

1

No details or no systematic plan

11