Product for advanced cell therapy

Abstract

The present invention relates to the development of a process for obtaining a product for advanced cell therapy capable of immunologically supporting patients.

Description

[0001] "Product for advanced cell therapy"

[0002] DESCRIPTION

[0003] The present invention relates to an advanced cell therapy product (ATMP ) for the immunological support of patients undergoing hematopoietic stem cell transplantation .

[0004] The clinical need arises from the fact that these patients are at risk of leukemic relapses and viral infections in the first three months after transplantation, a situation due to profound immunosuppression caused by pre-transplant ablative conditioning .

[0005] Cell therapy products are currently being developed that comprise only T lymphocytesyS or a speci fic subset ( in the absence of NK cells ) , but no processes are reported from which a cellular drug with the combination of T lymphocytes y5 and NK cells is obtained .

[0006] The prior art document EP 4 . 183871 describes the preparation of a cellular product lacking the V51 subset , while the amount of residual B lymphocytes is not described; the preparation protocol involves treatment with zoledronic acid for at least 5 days , Tap depletion prior to the culture phase , the use of animal FCS and does not involve medium changes with re-addition of cytokines .

[0007] The prior art document WO 2023 / 250336 describes the preparation of a cellular product that does not include NK by means of a speci fic expansion protocol , reagents and reagent concentration .

[0008] The prior art document CN 110564683 describes a 17 -20 day method, which employs non-clini cal grade products (RPMI ) , which employs antibodies ( anti-CD3 , anti-CD28 ) , employs multiple cytokines together ( IL2 , IL15 and IL21 ) and the addition of zoledronic acid in multiple steps ; the protocol does not provide for the depletion of Tap lymphocytes .

[0009] The prior art document CN 110283785 describes a method involving an initial separation and puri fication of Tyd and NK cells followed by co-cultivation .

[0010] The publication by Mohanadh Nada et al . ("Enhancing adoptive cancer immunotherapy with V [ gamma ] v2 [ delta ] T cells through pulse zoledronate stimulation" Journal for immunotherapy of cancer, Biomed Central Ltd, vol . 5 , no . 1 , 21 February 2017 , pages 1 -23 describes a method employing a di f ferent medium (RPMI or Optimi zer ) with animal-derived reagents and Tyd puri fication at the end of expansion .

[0011] The prior art document US 2020 / 032211 describes a method that includes the use of polypeptides and reagents and includes positive or negative puri fications of Tyd and / or NK; it does not employ human AB serum and does not provide for changes of medium with the addition of cytokines .

[0012] Summary of the invention

[0013] The inventors of the present patent application have surprisingly developed a process for obtaining an advanced cell therapy product (ATMP ) composed of innate immunity killer cells composed of expanded gamma delta T lymphocytes and Natural Killer cells .

[0014] Description of the figures

[0015] Figure 1 shows the plots obtained from the cytof luorimetric analysis of the cell suspension in the process steps . Panel A: MNC analysis after Ficoll gradient and before stimulation with ZOL and IL-2 . Plot a ) physical analysis of the cell population, b ) analysis of monocytes (CD14 positive cells, highlighted in the dashed box) necessary for the stimulation of T lymphocytesyS, c) presence of T lymphocytes (CD3 positive cells, highlighted in the dashed box) , d) presence of Ty8 and T lymphocytesaP (highlighted in the boxes) , e) analysis of the V5 1 and V6 2 subsets. Panel B: cytof luorimetric analysis showing the presence of TyS and T lymphocytesaP (highlighted in the boxes of the plot on the left) and of the V6 1 and V6 2 subsets (highlighted in the plot on the right) at the end of cell expansion, before the depletion of T lymphocytesap. Panel C: analysis, by cytof luorimetry, of TyS and T lymphocytesaP (highlighted in the boxes of the plot on the left) and of the V6 1 and V6 2 subsets (highlighted in the central plot) present in the cellular product after the depletion of T lymphocytesaP, which are in fact absent. The presence of NK cells is shown in the plot on the right and highlighted in the box (CD3 negative, CD56 positive cells) .

[0016] Panel A of Figure 2 shows: cell viability analysis (7-AAD negative cells, highlighted by the box) before (plot on the left) and after cryopreservation of the product (plot on the right) . Panel B of Figure 2 shows: analysis of the percentages of T lymphocytesyS and in panel C those of NK cells (plot on the left before freezing, plot on the right after thawing) .

[0017] In Figure 3, the top panels show the surface expression of CD107a on T lymphocytesyS (left) and NK cells (right) after culturing the cellular product in the presence of the K652 leukemic line; the bottom panels show similar results obtained after culturing with the SK-N-AS neuroblastoma line.

[0018] Object of the invention In a first object, the present invention describes a method for the preparation of an advanced cell therapy product.

[0019] In a second object, the present invention describes the advanced cell therapy product obtained with the described method.

[0020] In a third object, the present invention describes the advanced cell therapy product described for medical use.

[0021] Specific medical uses are further objects of the present invention .

[0022] In a further object, a method for treating an immunodeficiency state is described, comprising the step of administering to a patient in need a sufficient amount of the therapeutic product obtainable by the method of the present invention.

[0023] Detailed description of the invention

[0024] According to a first object, the present invention describes a method for the preparation of a therapeutic product.

[0025] In particular, said method comprises the steps of:

[0026] I) stimulation of mononuclear cells (MNC) ,

[0027] II) cell cultivation, phase III) depletion of Tap lymphocytes.

[0028] For the purposes of the present invention, the stimulation phase I) comprises the stimulation of mononuclear cells with human AB serum, zoledronic acid and interleukin 2 (IL-2) .

[0029] In particular, the cells are in a concentration of about 1-10 X106 / ml and preferably of about 5X106 / ml.

[0030] In particular, the stimulation with human AB serum employs human

[0031] AB serum in a concentration of about 4-20% and preferably 5%. In particular, stimulation with zoledronic acid employs zoledronic acid in a concentration of about 5-100 pM and preferably of about 25 pM.

[0032] In particular, the stimulation with interleukin 2 employs interleukin 2 (IL-2) in a concentration of about 10-2000 lU / ml and preferably of about 1000 lU / ml.

[0033] More specifically, phase II) of cell cultivation includes the sub-phases of: phase Ila) in which a change of culture medium is carried out phase lib) repetition of phase Ila) .

[0034] For the purposes of this invention, phase Ila) involves resuspending the cells in human AB serum from 4 to 20%, and preferably 5%, at the final concentration of about 1-10 X106 / ml and preferably about 2X106 / ml and then adding interleukin 2 (IL-2) in a concentration of 10-2000 lU / ml, and preferably in a concentration of 1000 lU / ml, and then incubating the cells at 37°C in the presence of 5% CO2.

[0035] For the purposes of the present invention, step lib) comprises repeating step Ila) on day 8° and day 11°.

[0036] According to one aspect of the present invention, in the described method, said step I) is carried out on isolated allogeneic or autologous mononuclear cells or obtained from a haploidentical donor .

[0037] According to one aspect of the present invention, in the method described after step I) of stimulation, said cells are incubated at 37°C in the presence of 5% CO2 for a period of 4 days. According to one aspect of the present invention, in the described method, said step III) is performed on the 14th day after the expansion phase.

[0038] According to one aspect of the present invention, in the described method, said step III) is performed up to a concentration of Tap lymphocytes lower than about 5%.

[0039] According to one aspect of the present invention, the described method comprises the further step of subjecting the cell preparation to centrifugation at 400 g for a time of about 10 minutes and at a temperature of about 19-25°C and elimination of the supernatant.

[0040] According to one aspect of the present, the method described comprises the further step of cryopreserving the cell preparation thus obtained.

[0041] According to a second object, the present invention describes the therapeutic product obtainable according to the above method.

[0042] In a preferred aspect of the present invention, the therapeutic product is characterized by comprising Tyd lymphocytes and NK cells.

[0043] More specifically, the therapeutic product of the invention is characterized by comprising:

[0044] Tyd lymphocytes >20%

[0045] NK cells >20%.

[0046] In a particularly preferred aspect of the present invention, the therapeutic product is characterized by comprising:

[0047] Tap lymphocytes <5% or even <1%

[0048] B lymphocytes <5% or even <1%.

[0049] In particular, in the therapeutic product of the present invention, the Tyd lymphocytes comprise: Tyd V51 lymphocytes about 1-30%

[0050] Tyd Vd2 lymphocytes about 60- 99% .

[0051] According to a third obj ect , the therapeutic product of the invention is described for medical use .

[0052] In particular, the therapeutic product of the invention is described for medical use in the treatment of an immunodeficiency state .

[0053] In one aspect of the invention, the therapeutic product of the invention is described for use in the prevention of viral infections .

[0054] In one aspect of the invention, the therapeutic product of the invention is described for use in the prevention of tumor relapses .

[0055] In particular, such relapses may be leukemia or neuroblastoma .

[0056] In one aspect of the invention, the therapeutic product of the invention is described for use in a state of immunodeficiency following a hematopoietic stem cell transplant or an organ transplant .

[0057] In one aspect of the invention, the therapeutic product of the invention is described for use in an immunodeficient state caused by a pre-transplant ablative treatment .

[0058] In one aspect of the invention, the therapeutic product of the invention is described for medical use in an adult or pediatric sub ect .

[0059] In one aspect of the invention, the therapeutic product of the invention is described for medical use in a subj ect who is an oncological or non-oncological patient . In one aspect of the invention, the therapeutic product of the invention is described for medical use in combination therapy with monoclonal antibodies .

[0060] In a further obj ect , a method for treating an immunodeficiency state is described, comprising the step of administering to a patient in need a suf ficient amount of the therapeutic product obtainable by the method of the present invention .

[0061] The invention will be further described by the following experimental part .

[0062] Process example

[0063] The production process was performed using clini cal grade or GMP grade reagents and a closed system guaranteed by the use of :

[0064] 1 ) CliniMACS Prodigy instrument

[0065] 2 ) bags for cell culture ( gas-permeable ) as well as for the centri fugation of the cell suspension

[0066] Step

[0067] Day 1 . MNC separation and stimulation with zoledronic acid (ZOL) and IL-2

[0068] Reagents used

[0069] TexMACS™ GMP (Miltenyi Biotec )

[0070] Ficoll Paque ( Cytiva )

[0071] CliniMACS PBS / EDTA (Miltenyi Biotec )

[0072] Human albumin 20% infusible ( CSL Behring)

[0073] AB serum from donor

[0074] Zoledronic acid ( Tillomed)

[0075] IL-2 ( Proleukin from Novartis ) Procedure

[0076] The separation of MNCs from leukapheresis was carried out using Module 2 ( called density gradient ) of the standardi zed procedure by the company Miltenyi within the protocol called adherent cell cul ture .

[0077] - Under a sterile hood, the leukapheresis was trans ferred to a 600 ml bag using a sterile connector .

[0078] - 150 ml of Ficoll Paque were placed in a 150 ml bag

[0079] - A solution of CliniMACS PBS / EDTA with 5% human albumin was prepared in a 2-liter bag

[0080] - The initiali zation of the CliniMACS Prodigy was performed according to the manufacturer ' s instructions (Miltenyi company) and selecting the "Adherent Cell Culture" process

[0081] - The bags described above and the tubing set on the CliniMACS Prodigy were installed according to the instructions of the company Miltenyi

[0082] - The preset indications in the program provided by the Miltenyi company were followed

[0083] - The MNC cells were automatically collected by the CliniMACS Prodigy in a bag with TexMACS™ GMP

[0084] - I f the cells were at a concentration of less than 5xl 06 / ml , they were centri fuged at 400 g, 10 minutes with brake 3 ( 19-25 ° C ) and resuspended in TEXMACS™ GMP with 5% AB serum at the final concentration of 5xl 06 / ml . The cells were trans ferred into a permeable gas bag and then ZOL 25 p M and 1000 lU / ml of IL-2 were added . - I f the cells had a concentration greater than 5xl 06 / ml , they were brought to the final concentration of 5xl 06 / ml by adding TEXMACS™ GMP with 5% AB serum, trans ferred to a gas-permeable bag and added ZOL 25|i M and 1000 lU / ml of IL-2 .

[0085] - The MNCs were placed in an incubator at 37 ° C with 5% CO2 .

[0086] Day 5 . Change of culture medium and addition of IL-2

[0087] The cells were trans ferred into a bag and centrifuged at 400 g, 10 minutes with brake 3 ( 19-25 ° C ) . After removing most of the supernatant using a bag-squeezer, the cells were counted and resuspended in TEXMACS™ GMP with 5% AB serum at the final concentration of 2xl 06 / ml . The cells were trans ferred into a permeable gas bag and 1000 lU / ml of IL-2 was then added . The bag with the cell suspension was placed in an incubator at 37 ° C with 5% CO2 .

[0088] Day 8-9. Change of culture medium and addition of IL-2

[0089] The operations described in the previous point were repeated .

[0090] Day 11-12 . Change of culture medium and addition of IL-2

[0091] The operations described on day 5 were repeated .

[0092] Day 14 . First day of T lymphocyte depletionaP

[0093] The depletion of T ap lymphocytes at the end of the 14 -day culture was carried out with the program called LP-TCRab-19 validated by the company Miltenyi Biotec, following the indications of the normal scal e or large scal e module , depending on the number of cells present . The CliniMACS Prodigy instrument was initiali zed and set up according to the instructions provided by the company Miltenyi . The bag containing the cells was taken from the incubator, the contents trans ferred to one or more bags ( depending on the total volume) and centrifuged at 400g for 10 minutes, brake 3 at 19-15°C. After removing part of the supernatant, the cell count and phenotype were performed (to define the % of T lymphocytes 0.0 useful for setting the program on the Prodigy instrument) . After preparing the necessary solutions, initializing the instrument, and setting up the tubing set, all the bags were placed in the designated housings, according to the instructions of the company Miltenyi Biotec.

[0094] The values for the number of cells, the percentage of T lymphocytes 0.0, and the total volume were then set in the CliniMACS Prodigy instrument program. The cell separation process was then started with the end of the process scheduled for the following day.

[0095] Day 15. End of the production process

[0096] Once the T lymphocyte a0 depletion process was completed, the bag called the target cell fraction containing the cell suspension was taken and transferred to a new bag (150 or 600 ml depending on the volume) . Centrifugation was performed at 400 g for 10 minutes at 19- 25°C. After removing the supernatant, the cells (final product / drug) were transferred to CryoMACS® Freezing Bags, the remaining volume was calculated and an equal volume of the cryopreservation solution was added. The freezing process was then performed.

[0097] Results

[0098] Day 1.

[0099] Number of cells in leukapheresis : 5.23xl09

[0100] MNC concentration obtained after Ficoll gradient: 29.52xl06 / ml

[0101] Volume: 110 ml

[0102] Total MNC obtained: 3.24xl09 A cellular phenotype was performed by cytof luorimetry using a panel of f luorochrome-conj ugated antibodies that showed the presence of T cells y5 (2% of total lymphomonocyte cells) , monocytes (25.5%) and non-T cells (50.6%) .

[0103] The MNCs were brought to a concentration of 5xl06 / ml by adding 538 ml of TEXMACS™ GMP with 5% AB serum and transferred into a 1000 ml gas permeable differentiation bag (product code 170-076-404) . The stimuli (25 M ZOL and 1000 TU / ml IL-2) were then added. The bag with the MNC was then placed in an incubator at 37 °C, 5% CO2.

[0104] Day 5.

[0105] After removing the bag with the cells from the incubator, the suspended cells were transferred to a 600 ml bag and centrifuged at 400 g for 10 min at 20°C.

[0106] After removing most of the supernatant, the cells remained in 100 ml of TEXMACS™ GMP with 5% AB serum and were counted.

[0107] Cell concentration: 19.44xl06 / ml

[0108] Total cells: 1.94xl09

[0109] 870 ml of TEXMACS™ GMP with 5% AB serum was added and transferred to a 1000 ml gas-permeable differentiation bag. IL-2 was added at the final concentration of 1000 lU / ml and the bag was placed in the incubator at 37°C, 5% CO2.

[0110] Day 8.

[0111] The same operations as on day 5 were performed with the following differences .

[0112] Cell concentration: 16.8xl06 / ml

[0113] Total cells: 1.68xl09

[0114] Added 740 ml of TEXMACS™ GMP with 5% AB serum and 1000 lU / ml IL-2 Day 11.

[0115] The same operations as on day 5 were performed with the following differences .

[0116] Cell concentration: 16.98xl06 / ml

[0117] Total cells: 1.69xl09

[0118] Added 745 ml of TEXMACS™ GMP with 5% AB serum and 1000 TU / ml IL-2 Day 14.

[0119] After taking the bag from the incubator, the cell suspension was divided into two 600 ml bags and centrifuged at 400g, 10 minutes, brake 3 at 19-25°C. After removing part of the supernatant, the cell suspension was collected in a single bag, the volume was calculated, the phenotype was performed (to know the present share of T lymphocytes 0.0 necessary to set the parameters on the Prodigy instrument) and the cells were counted.

[0120] Cell count: 14.34 xl06 / ml

[0121] Volume: 240 ml

[0122] Total cells: 3.44xl09

[0123] % lymphocytes T a0 : 31.94

[0124] After preparing the necessary solutions, initializing the instrument, and setting up the tubing set, all the bags were placed in the designated slots, according to the instructions of the company Miltenyi Biotec.

[0125] The normal scale program was set by entering all the required data.

[0126] Day 15.

[0127] After the depletion process of the T lymphocytes a0, the bag called target cell fraction was taken, the volume was calculated and viability and phenotype analyses were performed to characterize the cells present in the cellular product.

[0128] The cell suspension was in saline solution with 0.5% human albumin.

[0129] Cell count: 2.54 xl06 / ml

[0130] Volume: 181 ml

[0131] Total cells: 0.46xl09

[0132] Viable cells: 97.3%

[0133] T lymphocytesyS : 27%

[0134] T y6 V6 1 lymphocytes: 2.9%

[0135] T y6 V6 2 lymphocytes: 23.8%

[0136] NK cells : 64.4%

[0137] T lymphocytesaP: 0.06%

[0138] B lymphocytes: 2.5%

[0139] Monocytes: 0.01%

[0140] The cellular product was transferred to a new 600 ml bag and centrifuged at 400 g for 10 minutes, brake 3 at 20°C. After removing the supernatant, the remaining volume was calculated and an equal volume of the cryopreservation solution was added. The cellular product was then transferred into 250 ml CryoMACS® Freezing Bags by placing 50 ml in each bag. Freezing was then performed using the "Shock freezing" rapid freezer.

[0141] From the above description, the advantages offered by the present invention will be immediately apparent to the expert.

[0142] In particular, the product described by the present invention:

[0143] - does not comprise non-human or impurity components,

[0144] - comprises the V51 subset with anti-viral activity, - has greater clinical safety for the concentration of B lymphocytes

[0145] <5% ,

[0146] - thanks to the presence of T yd lymphocytes and NK cells , it has a greater therapeutic activity, can be used in the post-transplant phase , even in pediatric patients and for non-oncological diseases ,

[0147] - comprises the V51 subset ,

[0148] - in general , it can be used : in immunodeficiencies , in the posttransplant period, in advance to avoid infections or relapses , for pediatric use .

[0149] The product of the present invention, due to its cellular composition, can provide important immunological support to patients who have received an allogeneic transplant . Patients who resort to transplantation can be adults or children and be af fected by both oncological and non-oncological diseases . The product consists of cells that have bridging functions between innate and adaptive immunity and perform killer activities by recogni zing target cells ( e . g . virus or tumor infected cell s ) in a non-MHC restricted manner . This function is of great interest in the clinical field as T lymphocytesyS and NK cells , which represent the cellular totality of our drug, do not induce graft versus host di sease ( GvHD) once infused into the patient . The product , which contains the two main T 75 subsets , namely V51 and Va, has all the biological characteristics to meet the expressed therapeutic needs . The product can be used to protect the pediatric patient and prevent tumor relapses and viral infections in the period following stem cell transplantation from a haploidentical donor . This use can be extended not only to patients receiving an allogeneic transplant (not necessarily haploidentical ) but also to all clinical situations in which the patient , adult or pediatric, is in a state of profound immunosuppression that is di f ficult to manage clinically .

[0150] The product , due to its innovative cellular composition, provides an answer to the problem of the complex management of the transplanted patient . The infusion of the drug in the patient undergoing a haploidentical transplant , or more generally allogeneic, can represent an important curative resource . The problem of post-transplant viral reactivations , GvHD, and tumor relapses is still dif ficult to manage clinically . These problems occur in patients who undergo a hematopoietic stem cell transplant , but also in those who undergo solid organ transplants , and posttransplant complications are often the cause of death .

[0151] The advantages of using the product are mani fold and concern : the possibility of preparing the drug starting from cells obtained from any donor, without the need to choose a donor among the many potential ones , which saves time and money . The product is a universal drug because , despite being cellular, there is no need for donor-recipient compatibility, the cryopreservation of the drug, its ready availability, its usefulness in preventing viral infections and relapses .

[0152] The product can be used in the phase that directly follows the transplant , which is the period of greatest immunosuppression in the patient ( about 90 days ) . The preventive aspect is crucial in that most of the infections that occur during this period are caused by latent viruses which, in fact , are reactivated precisely because of the poor immune defenses of the host and are often lethal . In current clinical practice , these infections are controlled and / or treated with multiple pharmacological cycles that are not always decisive .

[0153] The product of the invention can therefore make such treatments with antiviral drugs unnecessary, which are often expensive , and allow easier clinical management of the patient . A similar argument can be made on tumor relapses that not only impact the quality of li fe of patients , but that involve long hospitali zation and high costs related to drugs and / or additional transplants .

Claims

CLAIMS1. A method for the preparation of a therapeutic product, comprising the steps ofI) stimulation of mononuclear cells (MNC) in a concentration of about 1-10 X106 / ml with human AB serum at a concentration of about 4-20%, zoledronic acid in a concentration of 5-100 pM and interleukin 2 (IL-2) in a concentration of 10-2000 lU / ml,II) cell cultivation with human AB serum and interleukin 2 (IL- 2) , said phase comprising the sub-phases of: phase Ila) in which a change of culture medium is carried out by resuspending the cells in human AB serum at a concentration of about 4-20% at the final concentration of 1-10 X106 / ml and then adding interleukin 2 (IL-2) in a concentration of 10-2000 lU / ml and then incubating the cells at 37°C in the presence of 5% CO2; phase lib) repetition of phase Ila) on day 8° and day 11°; phase III) depletion of Tap lymphocytes.

2. The method according to the preceding claim, wherein said phase I) is performed on isolated allogeneic or autologous mononuclear cells (MNC) or obtained from a haploidentical donor.

3. The method according to any one of the preceding claims, wherein after the stimulation phase I) , said cells are incubated at 37°C in the presence of 5% CO2 for a period of 4 days.

4. The method according to any one of the preceding claims, wherein said phase III) is conducted on the 14th day after the stimulation phase.

5. The method according to any one of the preceding claims, wherein said step III) is carried out up to a concentration of Tap lymphocytes lower than about 5%.

6. The method according to any one of the preceding claims, comprising the further step of subjecting the cell preparation to centrifugation at 400 g for a time of about 10 minutes and at a temperature of about 19-25°C and elimination of the supernatant.

7. The method according to any one of the preceding claims, comprising the further step of cryopreserving the cell preparation thus obtained.

8. The therapeutic product obtainable according to the method of any one of the preceding claims.

9. The therapeutic product obtainable according to the method of any one of the preceding claims 1 to 7, characterized in that it comprises :Tyd lymphocytes > 20%NK cells > 20%.

10. The therapeutic product according to the preceding claim, characterized in that it comprises:Tap lymphocytes <5%B lymphocytes <5%.

11. The therapeutic product according to the previous claim 9 or 10, wherein said Tyd lymphocytes comprise:Tyd Vdl lymphocytes about 1-30%Tyd Vd2 lymphocytes about 60-99%.

12. The therapeutic product according to any one of the preceding claims 9 to 11 for medical use.

13. The therapeutic product according to any one of the preceding claims 9 to 12 for medical use in the treatment of an immunodeficiency state .14 . The therapeutic product according to any one of the preceding claims 9 to 13 for use in the prevention of viral infections .15 . The therapeutic product according to any one of the preceding claims 9 to 14 for use in the prevention of tumor relapse .

16. The therapeutic product according to any one of the preceding claims 9 to 15 , wherein said immunodeficiency state is subsequent to a hematopoietic stem cell transplant or an organ transplant .17 . The therapeutic product according to any one of the preceding claims 9 to 16 wherein said immunodef iciency state is caused by a pre-transplant ablative treatment .18 . The therapeutic product according to any one of the preceding claims 9 to 17 wherein said medical use is in an adult or pediatric subj ect .

19. The therapeutic product according to any one of the preceding claims 9 to 18 , wherein said medical use is in a subj ect who is an oncological or non-oncological patient .20 . The therapeutic product according to any one of the preceding claims 9 to 19 , wherein said medical use is in combination therapy with monoclonal antibodies .21 . A method for treating an immunodeficiency state comprising the step of administering to a patient in need thereof a suf ficientamount of a therapeutic product obtainable by the method of one of claims 1 to 7 .

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