The 34 references with contexts in paper A. Mudunov, A. Ahundov, M. Bolotin, T. Braunschweig, А. Мудунов, А. Ахундов, М. Болотин, Т. Брауншвейг (2018) “Новые результаты исследования плоскоклеточного рака головы и шеи в отношении классификации и системной терапии. Расширенный обзор // An update on head and neck squamous cell carcinoma in respect to classification and systemic therapy. Extended review” / spz:neicon:ogsh:y:2018:i:1:p:48-55

1
Aldalwg M.A.H., Brestovac B. Human Papillomavirus Associated Cancers of the Head and Neck: An Australian Perspective. Head Neck Pathol
Total in-text references: 1
  1. In-text reference with the coordinate start=5497
    Prefix
    This group of carcinomas, head and neck squamous cell carcinoma (HNSCC), show a wide range of incidences among the world, being more prevalent in some parts of South America and Asia, due to consumption of alcohol, bethel quid or cigarette smoking and, rising in recent years, infection by high-risk human papilloma virus (HPV) types, mostly HPV type 16 and 18
    Exact
    [1, 2]
    Suffix
    . The way of interacting of HPV high risk types and the host genome is based on expression of viral gene products, E6 and E7, leading to an uncontrolled cell proliferation by inactivation of p53 and rb (retinoblastoma-gene product) [3].

2
17;11(3):377–84. 2. Marur S., D'Souza G., Westra W.H., Forastiere A.A. HPV-associated head and neck cancer: a virus-related cancer epidemic. Lancet Oncol 2010;11(8):781–9. DOI: 10.1016/S1470-2045(10)70017-6.
Total in-text references: 1
  1. In-text reference with the coordinate start=5497
    Prefix
    This group of carcinomas, head and neck squamous cell carcinoma (HNSCC), show a wide range of incidences among the world, being more prevalent in some parts of South America and Asia, due to consumption of alcohol, bethel quid or cigarette smoking and, rising in recent years, infection by high-risk human papilloma virus (HPV) types, mostly HPV type 16 and 18
    Exact
    [1, 2]
    Suffix
    . The way of interacting of HPV high risk types and the host genome is based on expression of viral gene products, E6 and E7, leading to an uncontrolled cell proliferation by inactivation of p53 and rb (retinoblastoma-gene product) [3].

3
Брауншвейг Т., Левандровски А., Смеетс Д., Болотин М.В. Плоскоклеточный рак головы и шеи: новые перспективы лечения? Опухоли головы и шеи 2013;(3):4–10. [Braunschweig T., Lewandrowski A., Smeets D., Bolotin M.V. Squamous cell carcinoma of the head and neck: new avenues of treatment? Opukholi golovy i shei = Head and Neck Tumors 2013;(3):4–10. (In Russ.)]. DOI: 10.17650/2222-1468-2013-0-3-4-10.
Total in-text references: 1
  1. In-text reference with the coordinate start=5736
    Prefix
    The way of interacting of HPV high risk types and the host genome is based on expression of viral gene products, E6 and E7, leading to an uncontrolled cell proliferation by inactivation of p53 and rb (retinoblastoma-gene product)
    Exact
    [3]
    Suffix
    . HNSCCs are linked to high morbidity and mortality rates (up to 50 %) due to the spectrum of affected patients and high recurrence rates. While the incidence of HPV related HNSCC rise, non-HPV related HNSCC are decreasing [4, 5].

4
Forman D., de Martel C., Lacey C.J. et al. Global burden of human papillomavirus and related diseases. Vaccine 2012;30(5): F12–23. DOI: 10.1016/j.vaccine.2012.07.055.
Total in-text references: 1
  1. In-text reference with the coordinate start=5962
    Prefix
    HNSCCs are linked to high morbidity and mortality rates (up to 50 %) due to the spectrum of affected patients and high recurrence rates. While the incidence of HPV related HNSCC rise, non-HPV related HNSCC are decreasing
    Exact
    [4, 5]
    Suffix
    . In general, the age-standardization frequency of head and neck cancer worldwide is 8.1 per 100,000. In 2008, 550,319 new head and neck cancer cases were diagnosed worldwide, while in 2012 that number grew to 599,637 with Наряду с общепринятыми схемами конкурентного химиолучевого лечения, включая таргетную терапию цетуксимабом, в последние 2 года появился совершенно новый класс препаратов – п

5
Hammarstedt L., Lindquist D., Dahlstrand H. et al. Human papillomavirus as a risk factor for the increase in incidence of tonsillar cancer. Int J Cancer 2006;119(11): 2620–3. DOI: 10.1002/ijc.22177.
Total in-text references: 1
  1. In-text reference with the coordinate start=5962
    Prefix
    HNSCCs are linked to high morbidity and mortality rates (up to 50 %) due to the spectrum of affected patients and high recurrence rates. While the incidence of HPV related HNSCC rise, non-HPV related HNSCC are decreasing
    Exact
    [4, 5]
    Suffix
    . In general, the age-standardization frequency of head and neck cancer worldwide is 8.1 per 100,000. In 2008, 550,319 new head and neck cancer cases were diagnosed worldwide, while in 2012 that number grew to 599,637 with Наряду с общепринятыми схемами конкурентного химиолучевого лечения, включая таргетную терапию цетуксимабом, в последние 2 года появился совершенно новый класс препаратов – п

6
Ferlay J., Soerjomataram I., Dikshit R. et al. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer 2015; 136(5):E359–86. DOI: 10.1002/ijc.29210.
Total in-text references: 1
  1. In-text reference with the coordinate start=9391
    Prefix
    рак головы и шеи, радио- и химиотерапия, ингибирование контрольных точек, ипилимумаб, ниволумаб, пембролизумаб, атезолизумаб, дурвалумаб Для цитирования: Мудунов A., Ахундов A., Болотин M., Брауншвейг T. Новые результаты исследования плоскоклеточного рака головы и шеи в отношении классификации и системной терапии. Расширенный обзор. Опухоли головы и шеи 2018;8(1):48–55. 324,834 related deaths
    Exact
    [6]
    Suffix
    . Classical site for HPV related squamous cell carcinoma (SCC) is the oropharynx, as palatinal or lingual tonsils, but lately, the number of HPV positively tested cases of sinonasal SCC and laryngeal SCC are rising or diagnosed in a significant number as well [7, 8].

7
Bishop J.A., Guo T.W., Smith D.F. et al. Human papillomavirus-related carcinomas of the sinonasal tract. Am J Surg Pathol 2013;37(2):185–92. DOI: 10.1097/ PAS.0b013e3182698673.
Total in-text references: 1
  1. In-text reference with the coordinate start=9655
    Prefix
    Classical site for HPV related squamous cell carcinoma (SCC) is the oropharynx, as palatinal or lingual tonsils, but lately, the number of HPV positively tested cases of sinonasal SCC and laryngeal SCC are rising or diagnosed in a significant number as well
    Exact
    [7, 8]
    Suffix
    . The 5-year survival rate is more favorable in HPV positive cases than in HPV negative cases, due to less recurrence rates and better response to radiochemotherapy [8, 9]. One explanation of better prognosis in HPV positive SCC is the intrinsic immune response to the tumor, reflected by infiltrating T-cells [10].

8
Shaughnessy J.N., Farghaly H., Wilson L. et al. HPV: a factor in organ preservation for locally advanced larynx and hypopharynx cancer? Am J Otolaryngol 2014;35(1):19–24. DOI: 10.1016/j.amjoto. 2013.08.006.
Total in-text references: 2
  1. In-text reference with the coordinate start=9655
    Prefix
    Classical site for HPV related squamous cell carcinoma (SCC) is the oropharynx, as palatinal or lingual tonsils, but lately, the number of HPV positively tested cases of sinonasal SCC and laryngeal SCC are rising or diagnosed in a significant number as well
    Exact
    [7, 8]
    Suffix
    . The 5-year survival rate is more favorable in HPV positive cases than in HPV negative cases, due to less recurrence rates and better response to radiochemotherapy [8, 9]. One explanation of better prognosis in HPV positive SCC is the intrinsic immune response to the tumor, reflected by infiltrating T-cells [10].

  2. In-text reference with the coordinate start=9826
    Prefix
    cell carcinoma (SCC) is the oropharynx, as palatinal or lingual tonsils, but lately, the number of HPV positively tested cases of sinonasal SCC and laryngeal SCC are rising or diagnosed in a significant number as well [7, 8]. The 5-year survival rate is more favorable in HPV positive cases than in HPV negative cases, due to less recurrence rates and better response to radiochemotherapy
    Exact
    [8, 9]
    Suffix
    . One explanation of better prognosis in HPV positive SCC is the intrinsic immune response to the tumor, reflected by infiltrating T-cells [10]. The classical surrogate marker for HPV positive cases is the immunohistochemical stain for p16 or the product of the gene CDKN2A [11].

9
Coordes A., Lenz K., Qian X. et al. Metaanalysis of survival in patients with HNSCC discriminates risk depending on combined HPV and p16 status. Eur Arch Otorhinolaryngol 2016;273(8):2157–69. DOI: 10.1007/s00405-015-3728-0.
Total in-text references: 2
  1. In-text reference with the coordinate start=9826
    Prefix
    cell carcinoma (SCC) is the oropharynx, as palatinal or lingual tonsils, but lately, the number of HPV positively tested cases of sinonasal SCC and laryngeal SCC are rising or diagnosed in a significant number as well [7, 8]. The 5-year survival rate is more favorable in HPV positive cases than in HPV negative cases, due to less recurrence rates and better response to radiochemotherapy
    Exact
    [8, 9]
    Suffix
    . One explanation of better prognosis in HPV positive SCC is the intrinsic immune response to the tumor, reflected by infiltrating T-cells [10]. The classical surrogate marker for HPV positive cases is the immunohistochemical stain for p16 or the product of the gene CDKN2A [11].

  2. In-text reference with the coordinate start=10818
    Prefix
    By publications using meta-analysis, it could be shown that p16 status is not a complete independent marker for outcome, but the combination of HPV testing by DNA extraction and PCR based detection: HPV+ / p16+ cases had the best prognosis, HPV– / p16+ intermediate and HPV+ / p16- and HPV– / p16– the worst
    Exact
    [9]
    Suffix
    . By screening HNSCC for HPV relation, p16 is the recommended marker, especially in oropharyngeal SCCs with a cut off value of 70 % stained tumor area [11]. There are no recommendations concerning the used antibody clone.

10
Badoual C., Hans S., Merillon N. et al. PD-1-expressing tumor-infiltrating T cells are a favorable prognostic biomarker in HPV-associated head and neck cancer. Cancer Res 2013;73(1):128–38. DOI: 10.1158/0008-5472.CAN-12-2606.
Total in-text references: 2
  1. In-text reference with the coordinate start=9972
    Prefix
    The 5-year survival rate is more favorable in HPV positive cases than in HPV negative cases, due to less recurrence rates and better response to radiochemotherapy [8, 9]. One explanation of better prognosis in HPV positive SCC is the intrinsic immune response to the tumor, reflected by infiltrating T-cells
    Exact
    [10]
    Suffix
    . The classical surrogate marker for HPV positive cases is the immunohistochemical stain for p16 or the product of the gene CDKN2A [11]. In oropharyngeal squamous cell carcinoma, in some extend depending on the country or continent, p16 marks HPV high-risk positive cases up to 100 %, but down to 80 % [12, 13], due to the prevalence of infections by subtype HPV type 16.

  2. In-text reference with the coordinate start=14435
    Prefix
    From beginning on, immunohistochemical expression of PD-L1 in tumor cells or so called “immune cells” in inflammatory infiltrates within the tumor (mainly histiocytic cell types) were chosen as a major depending variable in the analysis of efficacy of the drug [19]. The number of PD-1 positive lymphocytes within or adjacent to the tumor was discussed in the beginning but not followed
    Exact
    [10]
    Suffix
    . In the last years, an additional predictive biomarker for the positive response to checkpoint inhibitors revealed the tumor mutational burden, TMB. The TMB had been tested in HNSCC and showed to be much higher in non-HPV related carcinomas [20].

11
Lewis J.S. Jr., Beadle B., Bishop J.A. et al. Human papillomavirus testing in head and neck carcinomas: Guideline from the College of American Pathologists. Arch Pathol Lab Med 2017; Dec 18. DOI: 10.5858/ arpa.2017-0286-CP.
Total in-text references: 2
  1. In-text reference with the coordinate start=10108
    Prefix
    One explanation of better prognosis in HPV positive SCC is the intrinsic immune response to the tumor, reflected by infiltrating T-cells [10]. The classical surrogate marker for HPV positive cases is the immunohistochemical stain for p16 or the product of the gene CDKN2A
    Exact
    [11]
    Suffix
    . In oropharyngeal squamous cell carcinoma, in some extend depending on the country or continent, p16 marks HPV high-risk positive cases up to 100 %, but down to 80 % [12, 13], due to the prevalence of infections by subtype HPV type 16.

  2. In-text reference with the coordinate start=10970
    Prefix
    is not a complete independent marker for outcome, but the combination of HPV testing by DNA extraction and PCR based detection: HPV+ / p16+ cases had the best prognosis, HPV– / p16+ intermediate and HPV+ / p16- and HPV– / p16– the worst [9]. By screening HNSCC for HPV relation, p16 is the recommended marker, especially in oropharyngeal SCCs with a cut off value of 70 % stained tumor area
    Exact
    [11]
    Suffix
    . There are no recommendations concerning the used antibody clone. As it is well established as being strongly expressed in almost 100 % of SCC of the cervix uteri [14], it is up to the pathological laboratory to establish a specifically staining antibody using for example SCC of cervical SCC as a positive control.

12
Ndiaye C., Mena M., Alemany L. et al. HPV DNA, E6/E7 mRNA, and p16INK4a detection in head and neck cancers: a systematic review and meta-analysis. Lancet Oncol 2014;15(12):1319–31. DOI: 10.1016/S1470-2045(14)70471-1.
Total in-text references: 1
  1. In-text reference with the coordinate start=10278
    Prefix
    The classical surrogate marker for HPV positive cases is the immunohistochemical stain for p16 or the product of the gene CDKN2A [11]. In oropharyngeal squamous cell carcinoma, in some extend depending on the country or continent, p16 marks HPV high-risk positive cases up to 100 %, but down to 80 %
    Exact
    [12, 13]
    Suffix
    , due to the prevalence of infections by subtype HPV type 16. Other types, including type 18 does not seem to be related to stable p16 expression. By publications using meta-analysis, it could be shown that p16 status is not a complete independent marker for outcome, but the combination of HPV testing by DNA extraction and PCR based detection: HPV+ / p16+ cases had the best prognosis, HPV– /

13
Ang K.K., Harris J., Wheeler R. et al. Human papillomavirus and survival of patients with oropharyngeal cancer. N Engl J Med 2010;363(1):24–35. DOI: 10.1056/ NEJMoa0912217.
Total in-text references: 1
  1. In-text reference with the coordinate start=10278
    Prefix
    The classical surrogate marker for HPV positive cases is the immunohistochemical stain for p16 or the product of the gene CDKN2A [11]. In oropharyngeal squamous cell carcinoma, in some extend depending on the country or continent, p16 marks HPV high-risk positive cases up to 100 %, but down to 80 %
    Exact
    [12, 13]
    Suffix
    , due to the prevalence of infections by subtype HPV type 16. Other types, including type 18 does not seem to be related to stable p16 expression. By publications using meta-analysis, it could be shown that p16 status is not a complete independent marker for outcome, but the combination of HPV testing by DNA extraction and PCR based detection: HPV+ / p16+ cases had the best prognosis, HPV– /

14
Carozzi F., Gillio-Tos A., Confortini M. et al. Risk of high-grade cervical intraepithelial neoplasia during follow-up in HPVpositive women according to baseline p16INK4A results: a prospective analysis of a nested substudy of the NTCC randomised controlled trial. Lancet Oncol 2013;14(2):168–76. DOI: 10.1016/S14702045(12)70529-6.
Total in-text references: 1
  1. In-text reference with the coordinate start=11138
    Prefix
    By screening HNSCC for HPV relation, p16 is the recommended marker, especially in oropharyngeal SCCs with a cut off value of 70 % stained tumor area [11]. There are no recommendations concerning the used antibody clone. As it is well established as being strongly expressed in almost 100 % of SCC of the cervix uteri
    Exact
    [14]
    Suffix
    , it is up to the pathological laboratory to establish a specifically staining antibody using for example SCC of cervical SCC as a positive control. Diagnosis Diagnostic approaches include always a biopsy taken of the tumor side by endoscopy.

15
Muller S., Khuri F.R., Konoet S.A. et al. HPV positive squamous cell carcinoma of the oropharynx. Are we observing an unusual pattern of metastases? Head Neck Pathol 2012;6(3):336–44. DOI: 10.1007/s12105-012-0355-6. tests), also other commercially available antibodies developed for research use were tested. Here, a significant number of well working antibodies could be found [25] and (table 3). Thus, several alternative antibodies are available with much less costs for the pathologist and patient. Сonclusion Squamous cell carcinomas of the head and neck are since recent years a field of plasticity and new approaches: after research discovered and established the connection of HPV infection and a subpopulation of HNSCC, the upcoming drug group of checkpoint inhibitors is abo
Total in-text references: 1
  1. In-text reference with the coordinate start=11739
    Prefix
    Histologically, a squamous cell carcinoma can be challenging in case of low differentiation or basaloid differentiation and depending on the surrounding inflammatory infiltration. HPV positive squamous cell carcinomas show generally a distinct pattern of basaloid cell types and no typical squamous stratification or keratinized differentiated parts
    Exact
    [15]
    Suffix
    . As shown in figure 1, a typical keratinizing squamous cell carcinoma of the tongue, p16 negative, is shown in 1A and 1B. In comparison, a basaloid squamous cell carcinoma origin in the tonsil, p16 positive, is shown in 1C and 1D.

16
Adelstein D., Gillison M.L., Pfister D.G. et al. NCCN Guidelines Insights: Head and Neck Cancers, Version 2.2017. J Natl Compr Canc Netw 2017;15(6):761–70. DOI: 10.6004/jnccn.2017.0101.
Total in-text references: 2
  1. In-text reference with the coordinate start=12373
    Prefix
    In unresectable cases or unresectable recurrance, systemic therapy as a combinated chemoradiotherapy, using classical chemotherapeutics, for example cisplatin and radiation or targeted therapeutics, as cetuximab and radiation
    Exact
    [16]
    Suffix
    . In non-naso-pharyngeal SCC in cases of recurrence after platinum containing therapy or progression under platinum containing therapy, a relatively new therapeutic agent, Pembrolizumab (MSD), was approved by the FDA and EMA [16].

  2. In-text reference with the coordinate start=12603
    Prefix
    In non-naso-pharyngeal SCC in cases of recurrence after platinum containing therapy or progression under platinum containing therapy, a relatively new therapeutic agent, Pembrolizumab (MSD), was approved by the FDA and EMA
    Exact
    [16]
    Suffix
    . Pembrolizumab (MSD) counts to the new group of so called checkpoint-inhibitors, representing a class of therapeutics interacting with the communication of tumor cells and cytotoxic T-cells [17]. In general, checkpoint inhibitors are blocking the ligand binding of the tumor cell on the receptor on the cytotoxic T-cell in order to activate the T-cells to attack tumor cells.

17
Haddad R., Seiwert T., Pfister D.G. et al. Pembrolizumab after progression on platinum and cetuximab in head and neck squamous cell carcinoma (HNSCC): results from KEYNOTE-055. Ann Oncol 2016;27(6).
Total in-text references: 1
  1. In-text reference with the coordinate start=12799
    Prefix
    in cases of recurrence after platinum containing therapy or progression under platinum containing therapy, a relatively new therapeutic agent, Pembrolizumab (MSD), was approved by the FDA and EMA [16]. Pembrolizumab (MSD) counts to the new group of so called checkpoint-inhibitors, representing a class of therapeutics interacting with the communication of tumor cells and cytotoxic T-cells
    Exact
    [17]
    Suffix
    . In general, checkpoint inhibitors are blocking the ligand binding of the tumor cell on the receptor on the cytotoxic T-cell in order to activate the T-cells to attack tumor cells. In many different tumor entities, trials had been executed with different types of checkpoint inhibitors, mainly blocking interactions of PD-1 / PD-L1 and CD80 / CTLA4.

18
Barth C. Immunotherapy for Head and Neck Cancer: The Fourth Modality Has Arrived. Available at: http://www.ascopost. com/issues/march-10-2017/immunotherapy-for-head-and-neck-cancer-thefourth-modality-has-arrived/
Total in-text references: 2
  1. In-text reference with the coordinate start=13324
    Prefix
    In many different tumor entities, trials had been executed with different types of checkpoint inhibitors, mainly blocking interactions of PD-1 / PD-L1 and CD80 / CTLA4. By several authors and speakers, checkpoint inhibitors are called “the fourth modality” next to the well established treatment options: surgery, radiation and chemotherapy
    Exact
    [18]
    Suffix
    . One of the first tumor entities in which checkpoint inhibitors were tested in trials was the malignant melanoma of the skin. Here, CTLA4 inhibitor Ipilimumab (BMS) didn’t reveal high response rates (5–15 %), but still better than prior regimes.

  2. In-text reference with the coordinate start=13663
    Prefix
    Here, CTLA4 inhibitor Ipilimumab (BMS) didn’t reveal high response rates (5–15 %), but still better than prior regimes. With PD-1 inhibitors like Nivolumab, up to 30–45 % stable response rates were achievable
    Exact
    [18]
    Suffix
    . For HNSCC, in case of CTLA4 several inhibitors are under investigation and mostly in combined therapies with another drug, none of the studies revealed a predicting biomarker as a immunohistochemical staining [19].

19
Economopoulou P., Perisanidis C., Giotakis E.I., Psyrri A. The emerging role of immunotherapy in head and neck squamous cell carcinoma (HNSCC): anti-tumor immunity and clinical applications. Ann Transl Med 2016;4(9):173. DOI: 10.21037/atm.2016.03.34.
Total in-text references: 3
  1. In-text reference with the coordinate start=13877
    Prefix
    For HNSCC, in case of CTLA4 several inhibitors are under investigation and mostly in combined therapies with another drug, none of the studies revealed a predicting biomarker as a immunohistochemical staining
    Exact
    [19]
    Suffix
    . In case of PD-1 and PD-L1 inhibitors, many different drugs were developed (a selection is listed below) and set up in numerous trials from phase I to phase III. From beginning on, immunohistochemical expression of PD-L1 in tumor cells or so called “immune cells” in inflammatory infiltrates within the tumor (mainly histiocytic cell types) were chosen as a major depending variable in the anal

  2. In-text reference with the coordinate start=14308
    Prefix
    From beginning on, immunohistochemical expression of PD-L1 in tumor cells or so called “immune cells” in inflammatory infiltrates within the tumor (mainly histiocytic cell types) were chosen as a major depending variable in the analysis of efficacy of the drug
    Exact
    [19]
    Suffix
    . The number of PD-1 positive lymphocytes within or adjacent to the tumor was discussed in the beginning but not followed [10]. In the last years, an additional predictive biomarker for the positive response to checkpoint inhibitors revealed the tumor mutational burden, TMB.

  3. In-text reference with the coordinate start=19327
    Prefix
    situations, an immunohistological testing is useful to determine if the treatment with checkpoint inhibitors might work better: malignant melanoma and single treatment with Nivolumab due to higher side effects of Ipilimumab in combination therapy; or in patients who had to pay for the drug themselves. In general, there was no correlation mentioned concerning PD-L1 status and p16 / HPV
    Exact
    [19]
    Suffix
    . But HPV positivFig. 2. Cell interaction of checkpoint ligand/receptors: between antigenpresenting cells (APC) and T-cells via PD-1 and PD-L1 and PD-1 and PDL2 in order to inhibit cytotoxic effects on APC.

20
Chung C.H., Frampton G.M., Chalmers Z.R. et al. Genomic profiling of squamous malignancies across anatomic sites. J Clin Oncol 2017;35(15):11512. DOI: 10.1200/ JCO.2017.35.15_suppl.11512.
Total in-text references: 1
  1. In-text reference with the coordinate start=14684
    Prefix
    In the last years, an additional predictive biomarker for the positive response to checkpoint inhibitors revealed the tumor mutational burden, TMB. The TMB had been tested in HNSCC and showed to be much higher in non-HPV related carcinomas
    Exact
    [20]
    Suffix
    . In theory, TMB high tumors should be related to a high internal immune response due to a higher number of neo antigens on the cell surface. Other factors are discussed to play a role as microbiome or inflammatory signatures [21].

21
Blank C.U., Haanen J.B., Ribas A., Schumacher T.N. Cancer immunology. The “cancer immunogram”. Science 2016;352(6286):658–60.
Total in-text references: 1
  1. In-text reference with the coordinate start=14914
    Prefix
    In theory, TMB high tumors should be related to a high internal immune response due to a higher number of neo antigens on the cell surface. Other factors are discussed to play a role as microbiome or inflammatory signatures
    Exact
    [21]
    Suffix
    . As for Pembrolizumab (MSD), equally for Nivolumab (BMS), it inhibits the receptor on the T-cell (see figure 2). Other inhibiting drugs are blocking the ligand as there are Atezolizumab (Roche), Durvalumab (AstraZeneca) and Avelumab (Merck).

22
Thallinger C., Fureder T., Preusser M. et al. Review of cancer treatment with immune checkpoint inhibitors: Current concepts, expectations, limitations and pitfalls. Wien Klin Wochenschr 2018;130(3–4):85–91. DOI: 10.1007/ s00508-017-1285-9.
Total in-text references: 2
  1. In-text reference with the coordinate start=16139
    Prefix
    All histological photographs at ×200, using Hamamatsu Scanner, HT2.0 (Hamamatsu, Herrsching, Germany); immunohistochemical stain by monoclonal mouse anti-p16 by Merck (Merck, Darmstadt, Germany) using DAKO/Agilent Autostainer and Flex Kit (Agilent, Waldbronn, Germany) of the receptor programmed-cell death-1 (PD-1) and programmed cell death-ligand-1 (PD-L1) is meant to be suppressed
    Exact
    [22]
    Suffix
    . As depicted in figure 2, PD-L1 is not only expressed on tumor cells, but also on antigen-presenting cells, as dendritic cells. This kind of treatment is not meant to be curative but to control the disease in order to reduce tumor burden or to receive stable disease.

  2. In-text reference with the coordinate start=16539
    Prefix
    This kind of treatment is not meant to be curative but to control the disease in order to reduce tumor burden or to receive stable disease. Side-effects of these drugs are in general less severe, as inflammation of lungs, colon and liver, but mostly well treatable
    Exact
    [22]
    Suffix
    . So far, of the five mentioned actual drugs in use or in close future use, are primarily tested in second line and after successful first line application. Nivolumab and Pembrolizumab had been under investigation in Phase III trials in HNSCC: Nivolumab in CheckMate studies (No 141) and Pembrolizumab in Keynote studies (No 012, 048, 055).

23
Bahleda R., Braiteh F.S., Balmanoukian A.S. et al. Long-term safety and clinical outcomes of atezolizumab in head and neck cancer: phase la trial results. Ann Oncol 2017;28(5):v372–94. DOI: 0.1093/ annonc/mdx374.001a.
Total in-text references: 1
  1. In-text reference with the coordinate start=16976
    Prefix
    Nivolumab and Pembrolizumab had been under investigation in Phase III trials in HNSCC: Nivolumab in CheckMate studies (No 141) and Pembrolizumab in Keynote studies (No 012, 048, 055). Atezolizumab and Durvalumab had also been investigated on HNSCC in phase I and II studies
    Exact
    [23, 24]
    Suffix
    . In the trial of Atezolizumab, there could not be shown any connection of the response to HPV status (p16 positivity) or PD-L1 immunohistochemical expression. In Durvalumab, only tumors with a PD-L1 expression of at least 25 % of tumor cells were included.

24
Zandberg D., Algazi A., Jimeno A. et al. Durvalumab for recurrent/metastatic (R/M) head and neck squamous cell carcinoma (HNSCC): Preliminary results from a single-arm, phase 2 study. Ann Oncol 2017;28(5):v372–94. DOI: 0.1093/ annonc/mdx374.
Total in-text references: 2
  1. In-text reference with the coordinate start=16976
    Prefix
    Nivolumab and Pembrolizumab had been under investigation in Phase III trials in HNSCC: Nivolumab in CheckMate studies (No 141) and Pembrolizumab in Keynote studies (No 012, 048, 055). Atezolizumab and Durvalumab had also been investigated on HNSCC in phase I and II studies
    Exact
    [23, 24]
    Suffix
    . In the trial of Atezolizumab, there could not be shown any connection of the response to HPV status (p16 positivity) or PD-L1 immunohistochemical expression. In Durvalumab, only tumors with a PD-L1 expression of at least 25 % of tumor cells were included.

  2. In-text reference with the coordinate start=17304
    Prefix
    In the trial of Atezolizumab, there could not be shown any connection of the response to HPV status (p16 positivity) or PD-L1 immunohistochemical expression. In Durvalumab, only tumors with a PD-L1 expression of at least 25 % of tumor cells were included. Here p16, resp. HPV positive patients showed a better outcome
    Exact
    [24]
    Suffix
    . In actual studies, the single drug as second line or first line is no longer much under investigation, but combination with other drugs / therapies or an established treatment followed by a checkpoint inhibitor (see table 2).

25
Scheel A.H., Dietel M., Heukamp L.C. et al. Harmonized PD-L1 immunohistochemistry for pulmonary squamous-cell and adenocarcinomas. Mod Pathol 2016;29(10):1165–72. DOI: 10.1038/ modpathol.2016.117.
Total in-text references: 2
  1. In-text reference with the coordinate start=18447
    Prefix
    As demonstrated in figure 4 and revealed in harmonization studies of different groups, the antibody clones: 28.8, 22C3 and SP263 are staining more equally with differences in intensity. The clone SP142 stained significantly less tumor cells but highlight more immune cells
    Exact
    [25, 26]
    Suffix
    . Even if many trials resulted in an approval of one of the drugs in different entities without necessary immunohistochemical testing for PD-L1, it still remains a useful biomarker. In many entities, higher response rates or progression free survival time could be described in tumors showing high expression of PD-L1 in tumor cells.

  2. In-text reference with the coordinate start=23612
    Prefix
    Harmonization studies of different working groups, the antibody clones: 28.8, 22C3 and SP263 are staining more equally with differences in intensity. The clone SP142 stained significantly less tumor cells but highlight more immune cells
    Exact
    [25, 26]
    Suffix
    (see figure 3). Next, cut-off values of 1 % are extremely uneasy to read in histological section with a high proportion of tumor tissue. Therefore, Agilent / Dako and MSD organized training work-shops for pathologists.

26
Hirsch F.R., McElhinny A., Stanforth D. et al. PD-L1 immunohistochemistry assays for lung cancer: results from phase 1 of the Blueprint PD-L1 IHC Assay Comparison Project. J Thorac Oncol 2017;12(2):208– 22. DOI: 10.1016/j.jtho.2016.11.2228.
Total in-text references: 2
  1. In-text reference with the coordinate start=18447
    Prefix
    As demonstrated in figure 4 and revealed in harmonization studies of different groups, the antibody clones: 28.8, 22C3 and SP263 are staining more equally with differences in intensity. The clone SP142 stained significantly less tumor cells but highlight more immune cells
    Exact
    [25, 26]
    Suffix
    . Even if many trials resulted in an approval of one of the drugs in different entities without necessary immunohistochemical testing for PD-L1, it still remains a useful biomarker. In many entities, higher response rates or progression free survival time could be described in tumors showing high expression of PD-L1 in tumor cells.

  2. In-text reference with the coordinate start=23612
    Prefix
    Harmonization studies of different working groups, the antibody clones: 28.8, 22C3 and SP263 are staining more equally with differences in intensity. The clone SP142 stained significantly less tumor cells but highlight more immune cells
    Exact
    [25, 26]
    Suffix
    (see figure 3). Next, cut-off values of 1 % are extremely uneasy to read in histological section with a high proportion of tumor tissue. Therefore, Agilent / Dako and MSD organized training work-shops for pathologists.

27
Ferris R.L., Clump D.A., Ohr J. et al. Phase I trial of cetuximab, intensity modulated radiotherapy (IMRT), and ipilimumab in previously untreated, locally advanced head and neck squamous cell carcinoma (PULA HNSCC). Ann Oncol 2017;28(Suppl 5). DOI: 10.1093/annonc/ mdx374.014.
Total in-text references: 1
  1. In-text reference with the coordinate start=21913
    Prefix
    Actual studies on combination of checkpoint inhibitors and other treatments GroupDrugShort descriptionNameRef anti-CTLA4 Ipilimumab (BMS) Combination of Cetuximab, Radiation and Ipilimumab in first line PULA HNSCC–
    Exact
    [27]
    Suffix
    Ipilimumab plus NivolumabCheckMate651&714[28, 29] Tremelimumab (AstraZeneca)Tremelimumab plus DurvalumabKestrel[30] anti-PD-1 PembrolizumabCombination with Chemo and Radiation and Pembrolizumab–[31] Nivolumab Ipilimumab plus NivolumabCheckMate651&714[28, 29] Nivolumab plus ChemotherapyCheckmate 227[32] Induction of Chemo plus Nivolumab followed by surgery (similar to neoadjuvant)Optima2[33] anti

28
BMS. Study of Nivolumab in Combination With Ipilimumab Compared to the Standard of Care (Extreme Study Regimen) as First Line Treatment in Patients With Recurrent or Metastatic Squamous Cell Carcinoma of the Head and Neck (CheckMate 651). 2016. Available at: https://clinicaltrials.gov/ct2/show/ NCT02741570.
Total in-text references: 2
  1. In-text reference with the coordinate start=21958
    Prefix
    Actual studies on combination of checkpoint inhibitors and other treatments GroupDrugShort descriptionNameRef anti-CTLA4 Ipilimumab (BMS) Combination of Cetuximab, Radiation and Ipilimumab in first line PULA HNSCC–[27] Ipilimumab plus NivolumabCheckMate651&714
    Exact
    [28, 29]
    Suffix
    Tremelimumab (AstraZeneca)Tremelimumab plus DurvalumabKestrel[30] anti-PD-1 PembrolizumabCombination with Chemo and Radiation and Pembrolizumab–[31] Nivolumab Ipilimumab plus NivolumabCheckMate651&714[28, 29] Nivolumab plus ChemotherapyCheckmate 227[32] Induction of Chemo plus Nivolumab followed by surgery (similar to neoadjuvant)Optima2[33] anti-PD-L1DurvalumabDurvalumab plus TremelimumabKestre

  2. In-text reference with the coordinate start=22161
    Prefix
    GroupDrugShort descriptionNameRef anti-CTLA4 Ipilimumab (BMS) Combination of Cetuximab, Radiation and Ipilimumab in first line PULA HNSCC–[27] Ipilimumab plus NivolumabCheckMate651&714[28, 29] Tremelimumab (AstraZeneca)Tremelimumab plus DurvalumabKestrel[30] anti-PD-1 PembrolizumabCombination with Chemo and Radiation and Pembrolizumab–[31] Nivolumab Ipilimumab plus NivolumabCheckMate651&714
    Exact
    [28, 29]
    Suffix
    Nivolumab plus ChemotherapyCheckmate 227[32] Induction of Chemo plus Nivolumab followed by surgery (similar to neoadjuvant)Optima2[33] anti-PD-L1DurvalumabDurvalumab plus TremelimumabKestrel[30] Fig. 4.

29
BMS. Study of Nivolumab in Combination With Ipilimumab Versus Nivolumab in Combination With Ipilimumab Placebo in Patients With Recurrent or Metastatic Squamous Cell Carcinoma of the Head and Neck (CheckMate 714). 2018. Available at: https://clinicaltrials.gov/ct2/show/ NCT02823574.
Total in-text references: 2
  1. In-text reference with the coordinate start=21958
    Prefix
    Actual studies on combination of checkpoint inhibitors and other treatments GroupDrugShort descriptionNameRef anti-CTLA4 Ipilimumab (BMS) Combination of Cetuximab, Radiation and Ipilimumab in first line PULA HNSCC–[27] Ipilimumab plus NivolumabCheckMate651&714
    Exact
    [28, 29]
    Suffix
    Tremelimumab (AstraZeneca)Tremelimumab plus DurvalumabKestrel[30] anti-PD-1 PembrolizumabCombination with Chemo and Radiation and Pembrolizumab–[31] Nivolumab Ipilimumab plus NivolumabCheckMate651&714[28, 29] Nivolumab plus ChemotherapyCheckmate 227[32] Induction of Chemo plus Nivolumab followed by surgery (similar to neoadjuvant)Optima2[33] anti-PD-L1DurvalumabDurvalumab plus TremelimumabKestre

  2. In-text reference with the coordinate start=22161
    Prefix
    GroupDrugShort descriptionNameRef anti-CTLA4 Ipilimumab (BMS) Combination of Cetuximab, Radiation and Ipilimumab in first line PULA HNSCC–[27] Ipilimumab plus NivolumabCheckMate651&714[28, 29] Tremelimumab (AstraZeneca)Tremelimumab plus DurvalumabKestrel[30] anti-PD-1 PembrolizumabCombination with Chemo and Radiation and Pembrolizumab–[31] Nivolumab Ipilimumab plus NivolumabCheckMate651&714
    Exact
    [28, 29]
    Suffix
    Nivolumab plus ChemotherapyCheckmate 227[32] Induction of Chemo plus Nivolumab followed by surgery (similar to neoadjuvant)Optima2[33] anti-PD-L1DurvalumabDurvalumab plus TremelimumabKestrel[30] Fig. 4.

30
NN. AstraZeneca Head and Neck Trials. 27.10.2016. Available at: https:// www.astrazeneca.com/media-centre/ press-releases/2016/astrazeneca-headand-neck-cancer-trials-27102016.html.
Total in-text references: 2
  1. In-text reference with the coordinate start=22026
    Prefix
    Actual studies on combination of checkpoint inhibitors and other treatments GroupDrugShort descriptionNameRef anti-CTLA4 Ipilimumab (BMS) Combination of Cetuximab, Radiation and Ipilimumab in first line PULA HNSCC–[27] Ipilimumab plus NivolumabCheckMate651&714[28, 29] Tremelimumab (AstraZeneca)Tremelimumab plus DurvalumabKestrel
    Exact
    [30]
    Suffix
    anti-PD-1 PembrolizumabCombination with Chemo and Radiation and Pembrolizumab–[31] Nivolumab Ipilimumab plus NivolumabCheckMate651&714[28, 29] Nivolumab plus ChemotherapyCheckmate 227[32] Induction of Chemo plus Nivolumab followed by surgery (similar to neoadjuvant)Optima2[33] anti-PD-L1DurvalumabDurvalumab plus TremelimumabKestrel[30] Fig. 4.

  2. In-text reference with the coordinate start=22357
    Prefix
    (AstraZeneca)Tremelimumab plus DurvalumabKestrel[30] anti-PD-1 PembrolizumabCombination with Chemo and Radiation and Pembrolizumab–[31] Nivolumab Ipilimumab plus NivolumabCheckMate651&714[28, 29] Nivolumab plus ChemotherapyCheckmate 227[32] Induction of Chemo plus Nivolumab followed by surgery (similar to neoadjuvant)Optima2[33] anti-PD-L1DurvalumabDurvalumab plus TremelimumabKestrel
    Exact
    [30]
    Suffix
    Fig. 4. As demonstrated also in different studies, the three stains in A (clone 28.8), B (clone 22C3) and D (clone SP263) stain differently, but the same amount of tumor cells (staining counted irrespectively the strength).

31
MSD. Pembrolizumab in Combination With Cisplatin and Intensity Modulated Radiotherapy (IMRT) in Head and Neck Cancer. 2016. Available at: https:// clinicaltrials.gov/ct2/show/NCT02777385.
Total in-text references: 1
  1. In-text reference with the coordinate start=22107
    Prefix
    on combination of checkpoint inhibitors and other treatments GroupDrugShort descriptionNameRef anti-CTLA4 Ipilimumab (BMS) Combination of Cetuximab, Radiation and Ipilimumab in first line PULA HNSCC–[27] Ipilimumab plus NivolumabCheckMate651&714[28, 29] Tremelimumab (AstraZeneca)Tremelimumab plus DurvalumabKestrel[30] anti-PD-1 PembrolizumabCombination with Chemo and Radiation and Pembrolizumab–
    Exact
    [31]
    Suffix
    Nivolumab Ipilimumab plus NivolumabCheckMate651&714[28, 29] Nivolumab plus ChemotherapyCheckmate 227[32] Induction of Chemo plus Nivolumab followed by surgery (similar to neoadjuvant)Optima2[33] anti-PD-L1DurvalumabDurvalumab plus TremelimumabKestrel[30] Fig. 4.

32
BMS. An Investigational Immuno-therapy Trial of Nivolumab, or Nivolumab Plus Ipilimumab, or Nivolumab Plus Platinumdoublet Chemotherapy, Compared to Platinum Doublet Chemotherapy in Patients With Stage IV Non-Small Cell Lung Cancer (NSCLC) (CheckMate 227). 2015. Available at: https://clinicaltrials.gov/ct2/ show/NCT02477826.
Total in-text references: 1
  1. In-text reference with the coordinate start=22209
    Prefix
    Ipilimumab (BMS) Combination of Cetuximab, Radiation and Ipilimumab in first line PULA HNSCC–[27] Ipilimumab plus NivolumabCheckMate651&714[28, 29] Tremelimumab (AstraZeneca)Tremelimumab plus DurvalumabKestrel[30] anti-PD-1 PembrolizumabCombination with Chemo and Radiation and Pembrolizumab–[31] Nivolumab Ipilimumab plus NivolumabCheckMate651&714[28, 29] Nivolumab plus ChemotherapyCheckmate 227
    Exact
    [32]
    Suffix
    Induction of Chemo plus Nivolumab followed by surgery (similar to neoadjuvant)Optima2[33] anti-PD-L1DurvalumabDurvalumab plus TremelimumabKestrel[30] Fig. 4. As demonstrated also in different studies, the three stains in A (clone 28.8), B (clone 22C3) and D (clone SP263) stain differently, but the same amount of tumor cells (staining counted irrespectively the strength).

33
BMS. Oropharyngeal Tumor Induction Chemotherapy and Response-stratified Locoregional Therapy Trial in Order to Minimize Long-term Adverse Events (OPTIMA II). 2017. Available at: https:// clinicaltrials.gov/ct2/show/NCT03107182.
Total in-text references: 1
  1. In-text reference with the coordinate start=22298
    Prefix
    –[27] Ipilimumab plus NivolumabCheckMate651&714[28, 29] Tremelimumab (AstraZeneca)Tremelimumab plus DurvalumabKestrel[30] anti-PD-1 PembrolizumabCombination with Chemo and Radiation and Pembrolizumab–[31] Nivolumab Ipilimumab plus NivolumabCheckMate651&714[28, 29] Nivolumab plus ChemotherapyCheckmate 227[32] Induction of Chemo plus Nivolumab followed by surgery (similar to neoadjuvant)Optima2
    Exact
    [33]
    Suffix
    anti-PD-L1DurvalumabDurvalumab plus TremelimumabKestrel[30] Fig. 4. As demonstrated also in different studies, the three stains in A (clone 28.8), B (clone 22C3) and D (clone SP263) stain differently, but the same amount of tumor cells (staining counted irrespectively the strength).

34
Kintsler S. et al. PD-L1 Immunohistochemical stains and comparison of 10 available antibodies and their staining behaviour and costs. Mod Pathol, under Review 2018.
Total in-text references: 1
  1. In-text reference with the coordinate start=24044
    Prefix
    Due to high costs for the testing kits for the described antibodies (e. g. reagents only DAKO / Agilent: ~3150 € / 50 Table 3. Antibodies to test for PD-L1 immunohistochemistry (adjusted from S.Kintsler et al.
    Exact
    [34]
    Suffix
    ) CharacteristicAgilentAgilentVentanaVentanaCell SignallingAbcamZytomedQuartettMerck TypemMomRabmRabmRabmRabmMomRabmRabpolyclRab Clone28.822C3SP142SP263E1L3N28.8Cal10 QR1 Efficacy concordant++–+++++–