Figure 1

Figure 2

Cell numbers in both graphs are

difficult to compare because one
shows 10E9 and teh other 10E5-
10E8 …
I would say that cell growth with
IL-15RA is slower compared to
the presence of IL-2 over 3
weeks because you only reach
1x10E8 compared to 3x10E9
with IL-2 ??

Yes, the conclusion here is that

IL15-RA can support the growth,
however it is slower than with IL-
2. I am not sure with we can
directly compare both graphics.
Graphic A has Il-2 in the culture
media + Il-15 secreted by the
vectors…
Figure 3

Renata, Virginia, Killing of K562 by normal NK-92 is suboptimal at

30% and it leaves many questions. Why is killing of CAR.19
towards K562 reduced and restored when IL-15 is added ? My
explanation in this case would be NK-92 had not enough IL-2
thats why the overall killing is low and substitution of IL-15 in part
restored this by activating NK cells.

We dont know the answer, low Il-2 can be an explanation and

maybe IL-15 can interact/potentialize other NK receptors
Figure 4

Towards NK sensitive K562, which are devoid of CD19 there is highest release of IFN-Gamma,
TNF-alpha and IL-10, which is suprisingly highest in CAR NK cells carrying the IL-15 containing
CAR. In contrast to CD19 positive targets, where CAR-IL-15/IL-15RA yields highest releases of
above mentioned cytokines, CAR.19-IL-15/IL15RA shows background levels of IFN-gamma, TNF-
a and IL-10 in K562 cells.
This suggests that engagement of the CAR in CD19 positive Raji target cells activates intracellular
activation motifs which lead to enhanced cytokine production and release. Yes
better b/c DEG and not transcript

A C

Key DEG for

IL15/15RA:

E Integration and Summary:

B
Figure 6

For LAG – 3 the Y Achsis shows 1-5 % .. Basically there is no relevant activation
of LAG-3 no matter what. For PD-1, you see significant increase in expression
also in teh absence of target cells. It may be that the NK cells have not been
supplied well with IL-2 , which is why IL-15 containing CAR NK cells do overall
better … (intrinsic cytokine production) …
Figure 7

C
A

Tumor burden at day 29

Figure 9
Supplemental Figure 1